WorldWideScience

Sample records for genetic transformation

  1. Genetic Transformation in Citrus

    Directory of Open Access Journals (Sweden)

    Dicle Donmez

    2013-01-01

    Full Text Available Citrus is one of the world’s important fruit crops. Recently, citrus molecular genetics and biotechnology work have been accelerated in the world. Genetic transformation, a biotechnological tool, allows the release of improved cultivars with desirable characteristics in a shorter period of time and therefore may be useful in citrus breeding programs. Citrus transformation has now been achieved in a number of laboratories by various methods. Agrobacterium tumefaciens is used mainly in citrus transformation studies. Particle bombardment, electroporation, A. rhizogenes, and a new method called RNA interference are used in citrus transformation studies in addition to A. tumefaciens. In this review, we illustrate how different gene transformation methods can be employed in different citrus species.

  2. Genetic Transformation of Bacteria.

    Science.gov (United States)

    Moss, Robert.

    1991-01-01

    An activity in which students transform an ampicillin-sensitive strain of E. coli with a plasmid containing a gene for ampicillin resistance is described. The procedure for the preparation of competent cells and the transformation of competent E. coli is provided. (KR)

  3. Genetic Transformation of Switchgrass

    Science.gov (United States)

    Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

    Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

  4. Genetic Transformation of Streptococcus mutans

    OpenAIRE

    Perry, Dennis; Kuramitsu, Howard K.

    1981-01-01

    Three strains of Streptococcus mutans belonging to serotypes a, c, and f were transformed to streptomycin resistance by deoxyribonucleic acids derived from homologous and heterologous streptomycin-resistant strains of S. mutans and Streptococcus sanguis strain Challis. Homologous transformation of S. mutans was less efficient than heterologous transformation by deoxyribonucleic acids from other strains of S. mutans.

  5. Genetic transformation of carnation (Dianthus caryophylus L.).

    Science.gov (United States)

    Nontaswatsri, Chalermsri; Fukai, Seiichi

    2010-01-01

    This chapter describes a rapid and efficient protocol for explant preparation and genetic transformation of carnation. Node explants from greenhouse-grown plants and leaf explants from in vitro plants are infected with Agrobacterium tumefaciens AGL0 harboring pKT3 plasmid, consisting of GUS and NPTII genes. Explant preparation is an important factor to obtain the transformed plants. The GUS-staining area was located only on the cut end of explants and only explants with a cut end close to the connecting area between node and leaf, produced transformed shoots. The cocultivation medium is also an important factor for the successful genetic transformation of carnation node and leaf explants. High genetic transformation efficiency of node and leaf explants cocultured with Agrobacterium tumefaciens was achieved when the explants were cocultivated on a filter paper soaked with water or water and acetosyringone mixture (AS).

  6. Physical methods for genetic plant transformation

    Science.gov (United States)

    Rivera, Ana Leonor; Gómez-Lim, Miguel; Fernández, Francisco; Loske, Achim M.

    2012-09-01

    Production of transgenic plants is a routine process for many crop species. Transgenes are introduced into plants to confer novel traits such as improved nutritional qualities, tolerance to pollutants, resistance to pathogens and for studies of plant metabolism. Nowadays, it is possible to insert genes from plants evolutionary distant from the host plant, as well as from fungi, viruses, bacteria and even animals. Genetic transformation requires penetration of the transgene through the plant cell wall, facilitated by biological or physical methods. The objective of this article is to review the state of the art of the physical methods used for genetic plant transformation and to describe the basic physics behind them.

  7. Methods for genetic transformation in Dendrobium.

    Science.gov (United States)

    da Silva, Jaime A Teixeira; Dobránszki, Judit; Cardoso, Jean Carlos; Chandler, Stephen F; Zeng, Songjun

    2016-03-01

    The genetic transformation of Dendrobium orchids will allow for the introduction of novel colours, altered architecture and valuable traits such as abiotic and biotic stress tolerance. The orchid genus Dendrobium contains species that have both ornamental value and medicinal importance. There is thus interest in producing cultivars that have increased resistance to pests, novel horticultural characteristics such as novel flower colours, improved productivity, longer flower spikes, or longer post-harvest shelf-life. Tissue culture is used to establish clonal plants while in vitro flowering allows for the production of flowers or floral parts within a sterile environment, expanding the selection of explants that can be used for tissue culture or genetic transformation. The latter is potentially the most effective, rapid and practical way to introduce new agronomic traits into Dendrobium. Most (69.4 %) Dendrobium genetic transformation studies have used particle bombardment (biolistics) while 64 % have employed some form of Agrobacterium-mediated transformation. A singe study has explored ovary injection, but no studies exist on floral dip transformation. While most of these studies have involved the use of selector or reporter genes, there are now a handful of studies that have introduced genes for horticulturally important traits.

  8. Plant regeneration and genetic transformation in Jatropha

    KAUST Repository

    Sujatha, M.

    2012-07-01

    Jatropha curcas, a non-edible oil bearing species with multiple uses, and considerable economic potential is emerging as a potential biofuel plant. The limited knowledge of this species, low and inconsistent yields, the narrow genetic variability, and vulnerability to insects and diseases are major constraints in successful cultivation of Jatropha as a biofuel crop. Hence, genetic improvement of Jatropha is essential by conventional and modern biotechnological tools to use as a viable alternative source of bio-diesel. Realising its potential as a bio-energy crop, in vitro regeneration methods have been established to meet the demand of large scale supply of superior clones, and also as a prelude for genetic improvement of the species through transgenic approaches. In this chapter, an overview of in vitro tissue culture and genetic transformation of Jatropha is discussed. © 2013 Springer Science+Business Media Dordrecht. All rights are reserved.

  9. A Perspective on Hypericum perforatum Genetic Transformation.

    Science.gov (United States)

    Hou, Weina; Shakya, Preeti; Franklin, Gregory

    2016-01-01

    Hypericum perforatum (St John's wort) is a reservoir of diverse classes of biologically active and high value secondary metabolites, which captured the interest of both researchers and the pharmaceutical industry alike. Several studies and clinical trials have shown that H. perforatum extracts possess an astounding array of pharmacological properties. These properties include antidepressant, anti-inflammatory, antiviral, anti-cancer, and antibacterial activities; and are largely attributed to the naphtodianthrones and xanthones found in the genus. Hence, improving their production via genetic manipulation is an important strategy. In spite of the presence of contemporary genome editing tools, genetic improvement of this genus remains challenging without robust transformation methods in place. In the recent past, we found that H. perforatum remains recalcitrant to Agrobacterium tumefaciens mediated transformation partly due to the induction of plant defense responses coming into play. However, H. perforatum transformation is possible via a non-biological method, biolistic bombardment. Some research groups have observed the induction of hairy roots in H. perforatum after Agrobacterium rhizogenes co-cultivation. In this review, we aim at updating the available methods for regeneration and transformation of H. perforatum. In addition, we also propose a brief perspective on certain novel strategies to improve transformation efficiency in order to meet the demands of the pharmaceutical industry via metabolic engineering.

  10. Genetic transformation of the sugar beet plastome.

    Science.gov (United States)

    De Marchis, Francesca; Wang, Yongxin; Stevanato, Piergiorgio; Arcioni, Sergio; Bellucci, Michele

    2009-02-01

    It is very important for the application of chloroplast engineering to extend the range of species in which this technology can be achieved. Here, we describe the development of a chloroplast transformation system for the sugar beet (Beta vulgaris L. ssp. vulgaris, Sugar Beet Group) by biolistic bombardment of leaf petioles. Homoplasmic plastid-transformed plants of breeding line Z025 were obtained. Transformation was achieved using a vector that targets genes to the rrn16/rps12 intergenic region of the sugar beet plastome, employing the aadA gene as a selectable marker against spectinomycin and the gfp gene for visual screening of plastid transformants. gfp gene transcription and protein expression were shown in transplastomic plants. Detection of GFP in Comassie blue-stained gels suggested high GFP levels. Microscopy revealed GFP fluorescence within the chloroplasts. Our results demonstrate the feasibility of engineering the sugar beet chloroplast genome; this technology provides new opportunities for the genetic improvement of this crop and for social acceptance of genetically modified sugar beet plants.

  11. Genetic Programming Transforms in Linear Regression Situations

    Science.gov (United States)

    Castillo, Flor; Kordon, Arthur; Villa, Carlos

    The chapter summarizes the use of Genetic Programming (GP) inMultiple Linear Regression (MLR) to address multicollinearity and Lack of Fit (LOF). The basis of the proposed method is applying appropriate input transforms (model respecification) that deal with these issues while preserving the information content of the original variables. The transforms are selected from symbolic regression models with optimal trade-off between accuracy of prediction and expressional complexity, generated by multiobjective Pareto-front GP. The chapter includes a comparative study of the GP-generated transforms with Ridge Regression, a variant of ordinary Multiple Linear Regression, which has been a useful and commonly employed approach for reducing multicollinearity. The advantages of GP-generated model respecification are clearly defined and demonstrated. Some recommendations for transforms selection are given as well. The application benefits of the proposed approach are illustrated with a real industrial application in one of the broadest empirical modeling areas in manufacturing - robust inferential sensors. The chapter contributes to increasing the awareness of the potential of GP in statistical model building by MLR.

  12. Genetic transformation of Vitis vinifera via organogenesis.

    Science.gov (United States)

    Mezzetti, Bruno; Pandolfini, Tiziana; Navacchi, Oriano; Landi, Lucia

    2002-09-27

    Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. The current methods for the production of transgenic grape plants are based on Agrobacterium-mediated transformation followed by regeneration from embryogenic callus. However, grape embryogenic calli are laborious to establish and the phenotype of the regenerated plants can be altered. Transgenic grape plants (V. vinifera, table-grape cultivars Silcora and Thompson Seedless) were produced using a method based on regeneration via organogenesis. In vitro proliferating shoots were cultured in the presence of increasing concentrations of N6-benzyl adenine. The apical dome of the shoot was removed at each transplantation which, after three months, produced meristematic bulk tissue characterized by a strong capacity to differentiate adventitious shoots. Slices prepared from the meristematic bulk were used for Agrobacterium-mediated transformation of grape plants with the gene DefH9-iaaM. After rooting on kanamycin containing media and greenhouse acclimatization, transgenic plants were transferred to the field. At the end of the first year of field cultivation, DefH9-iaaM grape plants were phenotypically homogeneous and did not show any morphological alterations in vegetative growth. The expression of DefH9-iaaM gene was detected in transgenic flower buds of both cultivars. The phenotypic homogeneity of the regenerated plants highlights the validity of this method for both propagation and genetic transformation of table grape cultivars. Expression of the DefH9-iaaM gene takes place in young flower buds of transgenic plants from both grape cultivars.

  13. Regeneration and genetic transformation in the Vitis genus.

    NARCIS (Netherlands)

    Martinelli, L.

    1997-01-01

    This work is a contribution to the development of regeneration systems and genetic transformation in the Vitis genus and opens interesting perspectives to the application of molecular techniques for study interesting traits, as well as for genetic improvement of grape.Somatic embryogenesis from leaf

  14. Agrobacterium mediated genetic transformation and regeneration in ...

    African Journals Online (AJOL)

    Aghomotsegin

    2015-03-11

    Mar 11, 2015 ... Department of Genetic Engineering and Biotechnology, Faculty of ... The results of this study would be an effective tool for crop .... under same environment. ..... Verma (Eds.), Plant Molecular Biology Manual, 2nd ed., Kluwer.

  15. Plant regeneration and genetic transformation in switchgrass-A review

    Institute of Scientific and Technical Information of China (English)

    Paul Merrick; Shuizhang Fei

    2015-01-01

    Switchgrass is native to the tal grass prairie of North America. It is self-incompatible and has varied ploidy levels from diploid (2x) to dodecaploid (12x) with tetraploid and octoploid being the most common. The high yielding potential and the ability to grow wel in marginal lands make switchgrass an ideal species as a dedicated biomass producer for lignocel ulosic ethanol production. Genetic transformation is an important tool for studying gene function and for germplasm improvement in switchgrass, the genome of which has been sequenced recently. This paper intends to provide a comprehensive review on plant regeneration and genetic transformation in switchgrass. We ifrst reviewed the effect of explants, basal medium and plant growth regulators on plant regeneration in switchgrass, which is a prerequisite for genetic transformation. We then reviewed the progresses on genetic transformation with either the biolistic or Agrobacterium-mediated method in switchgrass, and discussed various techniques employed to improve the transformation efifciency. Final y we reviewed the recent progresses on the use of genetic transformation in improving biomass quality such as the reduction of lignin, and in increasing biomass yield in switchgrass. We also provided a future perspective on the use of new genome editing technologies in switchgrass and its potential impact on regulatory processes.

  16. Genetic Transformation System for the Fungal Soybean Pathogen Cercospora kikuchii.

    Science.gov (United States)

    Upchurch, R G; Ehrenshaft, M; Walker, D C; Sanders, L A

    1991-10-01

    An altered beta-tubulin gene that confers resistance to the fungicide benomyl was isolated from a genomic library of a UV-induced mutant of Cercospora kikuchii and used as a selectable marker for transformation. The level of benomyl resistance conferred to the transformants was at least 150-fold greater than the intrinsic resistance of the C. kikuchii recipient protoplasts. In the majority of cases, the tubulin fragment was integrated at the native beta-tubulin locus, apparently by gene replacement or gene conversion. The frequency of transformation ranged from 0.2 to 6 transformants per mug of DNA, depending on the recipient strain. Transformation with linearized plasmid resulted in a higher frequency, without changing the type of integration event. Transformants were phenotypically stable after eight consecutive transfers on medium without benomyl. This is the first report of a genetic transformation system for a Cercospora species.

  17. Genetic transformation of Eucalyptus camaldulensis by agrobalistic method

    Directory of Open Access Journals (Sweden)

    Evânia Galvão Mendonça

    2013-06-01

    Full Text Available Eucalyptus stands in the setting of worldwide forestry due to its adaptability, rapid growth, production of high-quality and low cost of wood pulp fibers. The eucalyptus convetional breeding is impaired mainlly by the long life cycle making the genetic transformation systems an important tool for this purpose. However, this system requires in vitro eficient protocols for plant induction, regeneration and seletion, that allow to obtain transgenic plants from the transformed cell groups. The aim of this work was to evaluate the callus formation and to optimize the leaves and callus genetic transformation protocol by using the Agrobacterium tumefaciens system. Concerning callus formation, two different culture media were evaluated: MS medium supplemented with auxin, cytokinin (M1 and the MS medium with reduced nitrogen concentration and supplemented with auxin, cytokinin coconut water (M2. To establish the leave genetic transformation, those were exposed to agrobiolistics technique (gene gun, to tissue injury, and A. tumesfasciens EHA 105 contening the vetor pCambia 3301 (35S::GUS::NOS, for gene transference and to establish the callus transformation thoses were exposed only to A. tumefasciens. For both experiments, the influence of different infection periods was evaluated. The M2 medium provided the best values for callus sizea and fresh and dry weight. The leaves genetic transformation using the agrobiolistics technique was effective, the gus gene transient expression could be observed. No significant differences were obtained in the infection periods (4, 6 and 8 minutes. The callus genetic transformation with A. tumefaciens also promotend the gus gene transient expression on the callus co-cultiveted for 15 e 30 minutes. The transformed callus was transfered to a regeneration and selection medium and transformed plants were obtained.

  18. Justice in the genetically transformed society.

    Science.gov (United States)

    Farrelly, Colin

    2005-03-01

    This paper explores some of the challenges raised by human genetic interventions for debates about distributive justice, focusing on the challenges that face prioritarian theories of justice and their relation to the argument advanced by Ronald Lindsay elsewhere in this issue of the Kennedy Institute of Ethics Journal. Also examined are the implications of germ-line genetic enhancements for intergenerational justice, and an argument is given against Fritz Allhoff's conclusion, found in this issue as well, that such enhancements are morally permissible if and only if they augment primary goods.

  19. Novel and potential application of cryopreservation to plant genetic transformation.

    Science.gov (United States)

    Wang, Biao; Zhang, Zhibo; Yin, Zhenfang; Feng, Chaohong; Wang, Qiaochun

    2012-01-01

    The world population now is 6.7 billion and is predicted to reach 9 billion by 2050. Such a rapid growing population has tremendously increased the challenge for food security. Obviously, it is impossible for traditional agriculture to ensure the food security, while plant biotechnology offers considerable potential to realize this goal. Over the last 15 years, great benefits have been brought to sustainable agriculture by commercial cultivation of genetically modified (GM) crops. Further development of new GM crops will with no doubt contribute to meeting the requirements for food by the increasing population. The present article provides updated comprehensive information on novel and potential application of cryopreservation to genetic transformation. The major progresses that have been achieved in this subject include (1), long-term storage of a large number of valuable plant genes, which offers a good potential for further development of novel cultivars by genetic transformation; (2), retention of regenerative capacity of embryogenic tissues and protoplasts, which ensures efficient plant regeneration system for genetic transformation; (3), improvement of transformation efficiency and plant regeneration of transformed cells; (4), long-term preservation of transgenic materials with stable expression of transgenes and productive ability of recombinant proteins, which allows transgenic materials to be stored in a safe manner before being analyzed and evaluated, and allows establishment of stable seed stocks for commercial production of homologous proteins. Data provided in this article clearly demonstrate that cryo-technique has an important role to play in the whole chain of genetic transformation. Further studies coupling cryotechnique and genetic transformation are expected to significantly improve development of new GM crops.

  20. Genetic transformation of plants by protoplast electroporation.

    Science.gov (United States)

    Bates, G W

    1994-10-01

    This article describes an optimized protocol for the electroporation of tobacco mesophyll protoplasts together with notes and data on the effects of various parameters and suggestions for work with protoplasts of other species. In this protocol, electroporation is achieved by means of electrical pulses from a high-voltage, capacitive-discharge unit. Procedures are described for measurement of protoplast viability with Evan's blue, the detection of transient expression of CAT and GUS gene plasmid constructs, and for the recovery of stable transformants based on selection for kanamycin resistance.

  1. The pathways of genetic transformation in cholangiocarcinogenesis.

    Science.gov (United States)

    Serafini, Francesco M; Radvinsky, David

    2016-12-01

    Cholangiocarcinoma (CCA) is an aggressive malignancy that originates from the epithelial cells of the biliary duct system. Depending on the anatomical location, CCA can be considered extrahepatic (eCCA) or intrahepatic (iCCA) (1). Two thirds of CCAs involve the extrahepatic biliary system, whereas the rest are confined within the liver parenchyma, beyond the secondary biliary radicals (2). Due to its biological aggressiveness and difficulty in diagnosis, the majority of patients with CCA are unresectable at presentation and the overall 5-year survival is approximately five percent (4). This article focuses on the genetic and epigenetic alterations present in cholangiocarcinomas, their occasional relationship to external stimuli, and with an emphasis on those unanswered questions about cholangiocarcinogenesis and future directions in the comprehension of epigenetic DNA methylation in patients with CCA.

  2. Agrobacterium-mediated genetic transformation of Camptotheca acuminata

    Institute of Scientific and Technical Information of China (English)

    WANG Hui-mei; ZU Yuan-gang

    2007-01-01

    UGPase gene related with wood cellulose synthesis was transferred into C. Acuminata using the method of Agrobacterium-mediated genetic transformation, and an efficient transformation system was developed for C. Acuminata on the basis of evaluations of several factors affecting Agrobacterium-mediated DNA transfer rate. The highest transformation rate was achieved when pre-cultured leaf explants were infected with an Agrobacterium culture corresponding to OD600 (0.5) for 10 min, and cultured on explant regeneration medium for three days. The results of Southern hybridization showed that genomic DNA of the kanamycin-resistant shoots to an UGPase gene probe substantiated the integration of the transgene. Transformation efficiency (6%) was achieved under the optimized transformation procedure. This system should facilitate the introduction of important useful genes into C. Acuminata.

  3. Agrobacterium rhizogenes Mediated Genetic transformation of Abrus precatorius L

    Directory of Open Access Journals (Sweden)

    Vijai Singh Karwasara

    2009-01-01

    Full Text Available Abrus precatorius L. known as Indian liquorice is a common deciduous vine containing sweet principle compound known as glycyrrhizin. Hence it can be used as a very good substitute for Liquorice. Genetic transformation has proved to be an effective way to enhance secondary metabolites in plant cell cultures. The transformation of Abrus precatorius L. mediated by Agrobacterium rhizogenes was studied using three bacterial strains i.e. MTCC 532, MTCCC 2364 and NCIM 5140. Putative hairy roots were obtained after the transformation. The effects of bacterial strains, bacterial concentration, acetosyringone and co-cultivation pH on transformation of Abrus were investigated. Co-cultivation with Strain MTCC 532 for 2 days with 100 µmol L -1 acetosyringone at pH 6.5 provided the optimal conditions under which transformation frequency approached 84%.

  4. Physical methods for genetic transformation of fungi and yeast

    Science.gov (United States)

    Rivera, Ana Leonor; Magaña-Ortíz, Denis; Gómez-Lim, Miguel; Fernández, Francisco; Loske, Achim M.

    2014-06-01

    The production of transgenic fungi is a routine process. Currently, it is possible to insert genes from other fungi, viruses, bacteria and even animals, albeit with low efficiency, into the genomes of a number of fungal species. Genetic transformation requires the penetration of the transgene through the fungal cell wall, a process that can be facilitated by biological or physical methods. Novel methodologies for the efficient introduction of specific genes and stronger promoters are needed to increase production levels. A possible solution to this problem is the recently discovered shock-wave-mediated transformation. The objective of this article is to review the state of the art of the physical methods used for genetic fungi transformation and to describe some of the basic physics and molecular biology behind them.

  5. Recent advances in the genetic transformation of coffee.

    Science.gov (United States)

    Mishra, M K; Slater, A

    2012-01-01

    Coffee is one of the most important plantation crops, grown in about 80 countries across the world. The genus Coffea comprises approximately 100 species of which only two species, that is, Coffea arabica (commonly known as arabica coffee) and Coffea canephora (known as robusta coffee), are commercially cultivated. Genetic improvement of coffee through traditional breeding is slow due to the perennial nature of the plant. Genetic transformation has tremendous potential in developing improved coffee varieties with desired agronomic traits, which are otherwise difficult to achieve through traditional breeding. During the last twenty years, significant progress has been made in coffee biotechnology, particularly in the area of transgenic technology. This paper provides a detailed account of the advances made in the genetic transformation of coffee and their potential applications.

  6. Recent Advances in the Genetic Transformation of Coffee

    Directory of Open Access Journals (Sweden)

    M. K. Mishra

    2012-01-01

    Full Text Available Coffee is one of the most important plantation crops, grown in about 80 countries across the world. The genus Coffea comprises approximately 100 species of which only two species, that is, Coffea arabica (commonly known as arabica coffee and Coffea canephora (known as robusta coffee, are commercially cultivated. Genetic improvement of coffee through traditional breeding is slow due to the perennial nature of the plant. Genetic transformation has tremendous potential in developing improved coffee varieties with desired agronomic traits, which are otherwise difficult to achieve through traditional breeding. During the last twenty years, significant progress has been made in coffee biotechnology, particularly in the area of transgenic technology. This paper provides a detailed account of the advances made in the genetic transformation of coffee and their potential applications.

  7. Genetic transformation of fruit trees: current status and remaining challenges.

    Science.gov (United States)

    Gambino, Giorgio; Gribaudo, Ivana

    2012-12-01

    Genetic transformation has emerged as a powerful tool for genetic improvement of fruit trees hindered by their reproductive biology and their high levels of heterozygosity. For years, genetic engineering of fruit trees has focussed principally on enhancing disease resistance (against viruses, fungi, and bacteria), although there are few examples of field cultivation and commercial application of these transgenic plants. In addition, over the years much work has been performed to enhance abiotic stress tolerance, to induce modifications of plant growth and habit, to produce marker-free transgenic plants and to improve fruit quality by modification of genes that are crucially important in the production of specific plant components. Recently, with the release of several genome sequences, studies of functional genomics are becoming increasingly important: by modification (overexpression or silencing) of genes involved in the production of specific plant components is possible to uncover regulatory mechanisms associated with the biosynthesis and catabolism of metabolites in plants. This review focuses on the main advances, in recent years, in genetic transformation of the most important species of fruit trees, devoting particular attention to functional genomics approaches and possible future challenges of genetic engineering for these species in the post-genomic era.

  8. Disentangling competence for genetic transformation and virulence in Streptococcus pneumoniae.

    Science.gov (United States)

    Lin, Jingjun; Zhu, Luchang; Lau, Gee W

    2016-02-01

    Horizontal gene transfer mediated by the competence regulon is a major driver of genome plasticity in Streptococcus pneumoniae. When pneumococcal cells enter the competent state, about 6% of the genes in the genome are up-regulated. Among these, some genes are essential for genetic transformation while others are dispensable for the process. Exhaustive deletion analyses show that some up-regulated genes dispensable for genetic transformation contribute to pneumococcal-mediated pneumonia and bacteremia infections. Interestingly, virulence functions of such genes are either dependent or independent of the competent state. Among the competent-state-dependent genes are those mediating allolysis, a process where small fraction of non-competent cells within the pneumococcal population are lysed by their competent counterparts, releasing DNA presumably for transformation. Inadvertently, the pore-forming toxin pneumolysin is also released during allolysis, contributing to virulence. In this review, we discuss recent advances in our understanding of pneumococcal virulence processes mediated by the competence regulon. We proposed that coupling of competence induction and bacterial fitness drives the natural selection to favor an intact competence regulon, which in turn, provides the long-term benefits of genetic plasticity.

  9. [PLANT GENETIC TRANSFORMATION USING CARBON NANOTUBES FOR DNA DELIVERY].

    Science.gov (United States)

    Burlaka, O M; Pirko, Ya V; Yemets, A I; Blume, Ya B

    2015-01-01

    The possibility of exploiting carbon nanotubes (CNTs)-based nanocarriers to deliver genes into protoplasts, callus and mesophyll explants of plants was examined. Using single-walled CNTs (SWCNTs) at the concentration of 20 μg/ml and multi-walled CNTs (MWCNTs) at the concentration of 15 μg/ml genetic transformation of Nicotiana tabacum L. mesophyll protoplasts with plasmid pGreen 0029 was carried out and transient expression of reporter yfp gene in the protoplasts was observed. Using SWCNTs at the concentration of 40 μg/ml and MWCNTs at the concentration of 30 μg/ml genetic transformation of N. tabacum callus and leaf explants with nptII gene as a part of plasmid pGreen 0029 was carried out. As a result plant regeneration on selective medium containing 50 mg/lkanamycin was shown. SWCNTs-based nanocarriers de-onstrated their appli-ability to transform protoplasts as well as walled plant cells. Whereas, MWCNTs-based nano-arriers were suitable only for transformation of proto-lasts due to the limiting role of cellulose walls in cell penetration.

  10. Constructing the barley model for genetic transformation in Triticeae

    Institute of Scientific and Technical Information of China (English)

    LÜ Bo; WU Jia-jie; FU Dao-lin

    2015-01-01

    Barley (Hordeum vulgare L.) is one of the oldest domesticated crops, showing dramatic adaptation to various climate and environmental conditions. As a major cereal crop, barley ranks the 4th after wheat, maize and rice in terms of planting area and production al over the world. Due to its diploid nature, the cultivated barley is considered as an ideal model to study the polyploid wheat and other Triticeae species. Here, we reviewed the development, optimization, and application of transgenic approaches in barley. The most efifcient and robust genetic transformation has been built on the Agrobacterium-mediated transfer in conjunction with the immature embryo-based regeneration. We then discussed future considerations of using more practical technologies in barley transformation, such as the T-DNA/transposon tagging and the genome editing. As a cereal crop amenable to genetic transformation, barley wil serve as the most valuable carrier for global functional genomics in Triticeae and is becoming the most practical model for generating value-added products.

  11. Cassava Genetic Transformation and its Application in Breeding

    Institute of Scientific and Technical Information of China (English)

    Jia Liu; Qijie Zheng; Qiuxiang Ma; Kranthi Kumar Gadidasu; Peng Zhang

    2011-01-01

    As a major source of food, cassava (Manihot esculenta Crantz) is an important root crop in the tropics and subtropics of Africa and Latin America, and serves as raw material for the production of starches and bioethanol in tropical Asia. Cassava improvement through genetic engineering not only overcomes the high heterozygosity and serious trait separation that occurs in its traditional breeding, but also quickly achieves improved target traits. Since the first report on genetic transformation in cassava in 1996, the technology has gradually matured over almost 15 years of development and has overcome cassava genotype constraints, changing from mode cultivars to farmer-preferred ones.Significant progress has been made in terms of an increased resistance to pests and diseases, biofortification, and improved starch quality,building on the fundamental knowledge and technologies related to planting, nutrition, and the processing of this important food crop that has often been neglected. Therefore, cassava has great potential in food security and bioenergy development worldwide.

  12. Genetic transformation of Pinus taeda by particle bombardment

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A protocol is presented for genetically engineering loblolly pine (Pinus taeda L.) using particle bombardment. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Plasmid pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) cryIAc coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (Nos) terminator sequences, and the selectable marker gene, neomycin phosphotransferase II (nptII) controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected by kanamycin resistance conferred by the introduced NPTII gene. Shoot regeneration was induced from the kanamycin-resistant callus, and transgenic plantlets were then produced. The presence of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern blot analysis, and insect feeding assays. The recovered transgenic plants were acclimatized and then established in soil.

  13. Biofuel Potential of Plants Transformed Genetically With NAC Family Genes

    Directory of Open Access Journals (Sweden)

    Sadhana eSingh

    2016-01-01

    Full Text Available NAC genes contribute to enhance survivability of plants under conditions of environmental stress and in secondary growth of the plants, thereby building biomass. Thus, genetic transformation of plants using NAC genes provides a possibility to tailor made biofuel plants. Over-expression studies have indicated that NAC family genes can provide tolerance to various biotic and abiotic stresses, either by physiological or biochemical changes at the cellular level, or by affecting visible morphological and anatomical changes, for example by development of lateral roots in a number of plants. Over-expression of these genes also work as triggers for development of secondary cell walls. In our laboratory, we have observed a NAC gene from Lepidium latifolium contributing to both enhanced biomass as well as cold stress tolerance of model plants tobacco. Thus, we have reviewed all the developments of genetic engineering using NAC genes which could enhance the traits required for biofuel plants, either by enhancing the stress tolerance or by enhancing the biomass of the plants. KeywordsNAC, Genetically engineered plants, Abiotic stress tolerance, Secondary growth, Cell wall synthesis, Biomass

  14. TRANSFORMATION

    Energy Technology Data Exchange (ETDEWEB)

    LACKS,S.A.

    2003-10-09

    Transformation, which alters the genetic makeup of an individual, is a concept that intrigues the human imagination. In Streptococcus pneumoniae such transformation was first demonstrated. Perhaps our fascination with genetics derived from our ancestors observing their own progeny, with its retention and assortment of parental traits, but such interest must have been accelerated after the dawn of agriculture. It was in pea plants that Gregor Mendel in the late 1800s examined inherited traits and found them to be determined by physical elements, or genes, passed from parents to progeny. In our day, the material basis of these genetic determinants was revealed to be DNA by the lowly bacteria, in particular, the pneumococcus. For this species, transformation by free DNA is a sexual process that enables cells to sport new combinations of genes and traits. Genetic transformation of the type found in S. pneumoniae occurs naturally in many species of bacteria (70), but, initially only a few other transformable species were found, namely, Haemophilus influenzae, Neisseria meningitides, Neisseria gonorrheae, and Bacillus subtilis (96). Natural transformation, which requires a set of genes evolved for the purpose, contrasts with artificial transformation, which is accomplished by shocking cells either electrically, as in electroporation, or by ionic and temperature shifts. Although such artificial treatments can introduce very small amounts of DNA into virtually any type of cell, the amounts introduced by natural transformation are a million-fold greater, and S. pneumoniae can take up as much as 10% of its cellular DNA content (40).

  15. Natural genetic transformation in Acinetobacter sp. BD413 Biofilms: introducing natural genetic transformation as a tool for bioenhancement of biofilm reactors

    Energy Technology Data Exchange (ETDEWEB)

    Hendrickx, L.

    2002-07-01

    This study focussed on the localization and quantification of natural genetic transformation using neutral and disadvantageous genes in monoculture biofilms to investigate gene transfer and expression of the transferred genes in the absence of a selective advantage. Data obtained by this investigation were regarded as initial steps for evaluating the applicability of adding catabolic traits into the indigenous bacterial community of biofilm reactors by in situ natural genetic transformation. Because Acinetobacter spp. strains are readily found in waste water treatment plants and because Acinetobacter sp. BD413 possesses a high effective level of competence, natural genetic transformation was investigated in monoculture Acinetobacter sp. BD413 biofilms. The genes used for transformation encoded for the green fluorescent protein (GFP) and its variants. Monitoring of transformation events were performed with the use of automated confocal laser scanning microscopy (CLSM) and semi automated digital image processing and analysis. (orig.)

  16. Aboveground insect infestation attenuates belowground Agrobacterium-mediated genetic transformation.

    Science.gov (United States)

    Song, Geun Cheol; Lee, Soohyun; Hong, Jaehwa; Choi, Hye Kyung; Hong, Gun Hyong; Bae, Dong-Won; Mysore, Kirankumar S; Park, Yong-Soon; Ryu, Choong-Min

    2015-07-01

    Agrobacterium tumefaciens causes crown gall disease. Although Agrobacterium can be popularly used for genetic engineering, the influence of aboveground insect infestation on Agrobacterium induced gall formation has not been investigated. Nicotiana benthamiana leaves were exposed to a sucking insect (whitefly) infestation and benzothiadiazole (BTH) for 7 d, and these exposed plants were inoculated with a tumorigenic Agrobacterium strain. We evaluated, both in planta and in vitro, how whitefly infestation affects crown gall disease. Whitefly-infested plants exhibited at least a two-fold reduction in gall formation on both stem and crown root. Silencing of isochorismate synthase 1 (ICS1), required for salicylic acid (SA) synthesis, compromised gall formation indicating an involvement of SA in whitefly-derived plant defence against Agrobacterium. Endogenous SA content was augmented in whitefly-infested plants upon Agrobacterium inoculation. In addition, SA concentration was three times higher in root exudates from whitefly-infested plants. As a consequence, Agrobacterium-mediated transformation of roots of whitefly-infested plants was clearly inhibited when compared to control plants. These results suggest that aboveground whitefly infestation elicits systemic defence responses throughout the plant. Our findings provide new insights into insect-mediated leaf-root intra-communication and a framework to understand interactions between three organisms: whitefly, N. benthamiana and Agrobacterium. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

  17. Advancing ecological understandings through technological transformations in noninvasive genetics

    Science.gov (United States)

    Albano Beja-Pereira; Rita Oliveira; Paulo C. Alves; Michael K. Schwartz; Gordon Luikart

    2009-01-01

    Noninvasive genetic approaches continue to improve studies in molecular ecology, conservation genetics and related disciplines such as forensics and epidemiology. Noninvasive sampling allows genetic studies without disturbing or even seeing the target individuals. Although noninvasive genetic sampling has been used for wildlife studies since the 1990s, technological...

  18. Improved genetic transformation of cork oak (Quercus suber L.).

    Science.gov (United States)

    Alvarez-Fernández, Rubén; Ordás, Ricardo-Javier

    2012-01-01

    An Agrobacterium-mediated transformation system for selected mature Quercus suber L. trees has been established. Leaf-derived somatic embryos in an early stage of development are inoculated with an AGL1 strain harboring a kanamycin-selectable plasmid carrying the gene of interest. The transformed embryos are induced to germinate and the plantlets transferred to soil. This protocol, from adult cork oak to transformed plantlet, can be completed in about one and a half years. Transformation efficiencies (i.e., percentage of inoculated explants that yield independent transgenic embryogenic lines) vary depending on the cork oak genotype, reaching up to 43%.

  19. Genetic changes in Mammalian cells transformed by helium cells

    Energy Technology Data Exchange (ETDEWEB)

    Durante, M.; Grossi, G. (Naples Univ. (Italy). Dipt. di Scienze Fisiche); Yang, T.C.; Roots, R. (Lawrence Berkeley Lab., CA (USA))

    1990-11-01

    Midterm Syrian Hamster embryo (SHE) cells were employed to study high LET-radiation induced tumorigenesis. Normal SHE cells (secondary passage) were irradiated with accelerated helium ions at an incident energy of 22 MeV/u (9--10 keV/{mu}m). Transformed clones were isolated after growth in soft agar of cells obtained from the foci of the initial monolayer plated postirradiation. To study the progression process of malignant transformation, the transformed clones were followed by monolayer subculturing for prolonged periods of time. Subsequently, neoplasia tests in nude mice were done. In this work, however, we have focused on karyotypic changes in the banding patterns of the chromosomes during the early part of the progressive process of cell transformation for helium ion-induced transformed cells. 26 refs., 5 figs., 2 tabs.

  20. Current status and trends of wheat genetic transformation studies in China

    Institute of Scientific and Technical Information of China (English)

    HE Yi; WANG Qiong; ZENG Jian; SUN Tao; YANG Guang-xiao; HE Guang-yuan

    2015-01-01

    More than 20 years have passed since the ifrst report on successful genetic transformation of wheat. With the establishment and improvement of transformation platform, great progresses have been made on wheat genetic transformation both on its fundamental and applied studies in China, especial y driven by the National Major Project for Transgenic Organism Breed-ing, China, initiated in 2008. In this review, wheat genetic transformation platform improvement and transgenic research progresses including new techniques applied and functional studies of wheat quality, yield and stress tolerant related genes and biosafety assessment are summarized. The existing problems and the trends in wheat transformation with traditional methods combined with genomic studies and genome editing technology are also discussed.

  1. Current status of genetic transformation technology developed in cucumber (Cucumis sativus L.)

    Institute of Scientific and Technical Information of China (English)

    WANG Shun-li; Seong Sub Ku; YE Xing-guo; HE Cong-fen; Suk Yoon Kwon; Pil Son Choi

    2015-01-01

    Genetic transformation is an important technique for functional genomics study and genetic improvement of plants. Until now, Agrobacterium-mediated transformation methods using cotyledon as explants has been the major approach for cu-cumber, and its frequency has been up to 23%. For example, signiifcantly enhancement of the transformation efifciency of this plant species was achieved from the cotyledon explants of the cultivar Poinsett 76 infected by Agrobacterium strains EHA105 with efifcient positive selection system in lots of experiments. This review is to summarize some key factors in-lfuencing cucumber regeneration and genetic transformation, including target genes, selection systems and the ways of transgene introduction, and then to put forward some strategies for the increasing of cucumber transformation efifciency. In the future, it is high possible for cucumber to be potential bioreactor to produce vaccine and biomaterials for human beings.

  2. Genetic transformation in two potato cultivars with T-DNA from disarmed Agrobacterium

    NARCIS (Netherlands)

    Ooms, G.; Burrell, M.M.; Karp, A.; Bevan, M.; Hille, J.

    1987-01-01

    Derivatives of potato (Solanum tuberosum cv.'s 'Maris Bard' and 'Desiree') transformed with disarmed T-DNA from genetically engineered Agrobacterium tumefaciens strains were isolated. The transformed plants were recovered from shoot-forming tumours induced by infection of wounds with mixed-cultures

  3. Efficient genetic transformation of red raspberry, Rubus ideaus L.

    Science.gov (United States)

    Mathews, H; Wagoner, W; Cohen, C; Kellogg, J; Bestwick, R

    1995-05-01

    We have developed an efficient transformation system for red raspberry (Rubus ideaus L.) using Agrobacterium mediated gene transfer. Using this system we have successfully introduced a gene that encodes an enzyme, S-adenosylmethionine hydrolase (SAMase), in raspberry cultivars Meeker (MK), Chilliwack (CH) and Canby (CY). Leaf and petiole expiants were inoculated with disarmed Agrobacterium tumefaciens strain EHA 105 carrying either of two binary vectors, pAG1452 or pAG1552, encoding gene sequences for SAMase under the control of the wound and fruit specific tomato E4 promoter. Primary shoot regenerants on selection medium were chimeral containing both transformed and non-transformed cells. Non-chimeral transgenic clones were developed by iterative culture of petiole, node and leaf explants, on selection medium, from successive generations of shoots derived from the primary regenerants. Percent recovery of transformants was higher with the selection marker gene hygromycin phosphotransferase (hpt), than with neomycin phosphotransferase (nptII). Transformation frequencies of 49.6%, 0.9% and 8.1% were obtained in cultivars Meeker, Chilliwack and Canby respectively from petiole expiants using hygromycin selection. Genomic integration of transgenes was confirmed by Southern hybridization. Transgenic plants from a total of 218 independent transformation events (161 MK, 4 CH, 53 CY) have been successfully established in soil.

  4. Combined genetic transformation and nutritional assay for identification of Moraxella nonliquefaciens.

    OpenAIRE

    1987-01-01

    A combined genetic transformation and nutritional assay is described that permits definitive identification of clinically isolated strains of Moraxella nonliquefaciens. Crude DNA preparations of strains of various Moraxella species were used to transform nutritional mutants of a stably competent strain of M. nonliquefaciens for ability to grow on a defined medium (Mn-B). DNA samples from 24 independently isolated strains of M. nonliquefaciens all resulted in massive (4+) transformation of eac...

  5. Regeneration and genetic transformation in the Vitis genus = [Regeneratie en genetische transformatie in het genus Vitis =

    NARCIS (Netherlands)

    Martinelli, L.

    1997-01-01


    This work is a contribution to the development of regeneration systems and genetic transformation in the Vitis genus and opens interesting perspectives to the application of molecular techniques for study interesting traits, as well as for genetic improvement of

  6. Morphological changes in atypical bird's foot trefoil plants obtained during genetic transformation by agrobacterium

    Directory of Open Access Journals (Sweden)

    Nikolić Radomirka R.

    2003-01-01

    Full Text Available Atypical plants of bird's foot trefoil (Lotus corniculatus L., Bokor cv showing altered morphological characters that deviate from a normal phenotype were found after plant regeneration from transformed tissue. It had been obtained by genetic transformation of root sections of seedlings using Agrobacterium tumefaciens vector LBA4404/pBI121 on a medium supplemented with 0.2 mg I-1 BAP. The transformants 2b arid 4a were found to have a greatly atypical habit, including shortened internodes, elongated leaves, regular leaf arrangement along the stem and thicker leaves. Inheritance of altered characters was observed in the first progeny generation, and their genetic origin was considered.

  7. Improved electroporation procedure for genetic transformation of Dekkera/Brettanomyces bruxellensis.

    Science.gov (United States)

    Miklenić, Marina; Žunar, Bojan; Štafa, Anamarija; Svetec, Ivan-Krešimir

    2015-12-01

    Yeast Dekkera/Brettanomyces bruxellensis is one of the most common contaminants in wine industry, but also one of the most promising candidates for large-scale bioethanol production. Brettanomyces bruxellensis not only produces and tolerates high ethanol concentrations, but can also ferment cellobiose and adapt to lignocellulose hydrolasate. Furthermore, genome sequences of several B. bruxellensis strains are available, and efforts have been made to develop tools for genetic transformation of this yeast. Previously, we reported a successful transformation using lithium acetate/PEG method and electroporation, however, with very low transformation efficiency (10-20 transformants μg(-1)). Here we describe an optimization of electroporation procedure which resulted in a significant increase of transformation efficiency (2.8 × 10(3) transformants μg(-1)). Several key transformation parameters were optimized including cell growth phase, density of cells in the transformation sample and electroporation settings. We determined that treating the cells with both lithium acetate (100 mM) and dithiothreitol (35 mM) synergistically improves transformation efficiency. Using the described procedure around 500 transformants can be obtained per transformation sample with 180 ng of non-homologous linear transforming fragment. Additionally, several transformants were obtained with less than 1 ng of DNA demonstrating that this procedure is adequate even when very limited amount of DNA is available. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Development of genetic transformation methodologies for an industrially-promising microalga: Scenedesmus almeriensis.

    Science.gov (United States)

    Dautor, Yasmeen; Úbeda-Mínguez, Patricia; Chileh, Tarik; García-Maroto, Federico; Alonso, Diego López

    2014-12-01

    The development of the microalgal industry requires advances in every aspect of microalgal biotechnology. In this regard, the availability of genetic engineering tools for industrially-promising species is key. As Scenedesmus almeriensis has promise for industrial use, we describe here an Agrobacterium-based methodology that allows stable genetic transformation of it for the first time, thus opening the way to its genetic manipulation. Transformation was accomplished using two different antibiotic resistance genes [hygromicine phophotransferase (hpt) and Shble] and it is credited by PCR amplification of both hpt/Shble and GUS genes and by the β-glucuronidase activity of transformed cells. Nevertheless, the single 35S promoter seems unable to direct gene expression to a convenient level in S. almeriensis as suggested by the low GUS enzymatic activity. Temperature was critical for the transformation efficiency.

  9. Genetic transformation of Metroxylon sagu (Rottb.) cultures via Agrobacterium-mediated and particle bombardment.

    Science.gov (United States)

    Ibrahim, Evra Raunie; Hossain, Md Anowar; Roslan, Hairul Azman

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.

  10. Genetic Transformation of Metroxylon sagu (Rottb. Cultures via Agrobacterium-Mediated and Particle Bombardment

    Directory of Open Access Journals (Sweden)

    Evra Raunie Ibrahim

    2014-01-01

    Full Text Available Sago palm (Metroxylon sagu is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L. Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.

  11. Natural genetic transformation generates a population of merodiploids in Streptococcus pneumoniae.

    Directory of Open Access Journals (Sweden)

    Calum Johnston

    Full Text Available Partial duplication of genetic material is prevalent in eukaryotes and provides potential for evolution of new traits. Prokaryotes, which are generally haploid in nature, can evolve new genes by partial chromosome duplication, known as merodiploidy. Little is known about merodiploid formation during genetic exchange processes, although merodiploids have been serendipitously observed in early studies of bacterial transformation. Natural bacterial transformation involves internalization of exogenous donor DNA and its subsequent integration into the recipient genome by homology. It contributes to the remarkable plasticity of the human pathogen Streptococcus pneumoniae through intra and interspecies genetic exchange. We report that lethal cassette transformation produced merodiploids possessing both intact and cassette-inactivated copies of the essential target gene, bordered by repeats (R corresponding to incomplete copies of IS861. We show that merodiploidy is transiently stimulated by transformation, and only requires uptake of a ~3-kb DNA fragment partly repeated in the chromosome. We propose and validate a model for merodiploid formation, providing evidence that tandem-duplication (TD formation involves unequal crossing-over resulting from alternative pairing and interchromatid integration of R. This unequal crossing-over produces a chromosome dimer, resolution of which generates a chromosome with the TD and an abortive chromosome lacking the duplicated region. We document occurrence of TDs ranging from ~100 to ~900 kb in size at various chromosomal locations, including by self-transformation (transformation with recipient chromosomal DNA. We show that self-transformation produces a population containing many different merodiploid cells. Merodiploidy provides opportunities for evolution of new genetic traits via alteration of duplicated genes, unrestricted by functional selective pressure. Transient stimulation of a varied population of

  12. Genetic transformation and phylogeny of bacterial symbionts from tsetse.

    Science.gov (United States)

    Beard, C B; O'Neill, S L; Mason, P; Mandelco, L; Woese, C R; Tesh, R B; Richards, F F; Aksoy, S

    1993-01-01

    Two isolates of bacterial endosymbionts, GP01 and GM02, were established in cell free medium from haemolymph of the tsetse, Glossina pallidipes and G. morsitans. These microorganisms appear similar to rickettsia-like organisms reported previously from various tsetse species. The 16S rRNA sequence analysis, however, placed them within the gamma subdivision of the Proteobacteria, phylogenetically distinct from most members of the Rickettsiaceae which align with the alpha subdivision. Distinct multiple endogenous plasmids are harboured by GP01 and GM02, suggesting that the two isolates are different. Restriction mapping analysis showed that one of the conserved plasmids is present in high copy number and is at least 80 kb in size. A heterologous plasmid pSUP204, which contains the broad host range oriV replication origin, was used to transfect bacterial cultures. The symbiont GM02 was transformed, and it expressed plasmid encoded resistance to the antibiotics ampicillin, tetracycline and chloramphenicol. Transformation of these symbionts may provide a novel means for expressing anti-parasitic genes within tsetse populations.

  13. Genetic transformation of Begonia tuberhybrida by Ri rol genes.

    Science.gov (United States)

    Kiyokawa, S; Kikuchi, Y; Kamada, H; Harada, H

    1996-04-01

    We have developed an Agrobacterium -mediated transformation system for commercial Begonia species. The leaf explants of Begonia semperflorens, Begonia x hiemalis and B. tuberhybrida were inoculated with Agrobacterium tumefaciens LBA4404 harboring a binary vector pBI121 which contains rolA, B and C genes of an agropine type Ri plasmid (pRiA4b). Kanamycin resistant shoots of B. tuberhybrida were obtained on MS agar medium supplemented with 0.1 mg/l NAA, 0.5 mg/l BA, 500 mg/l claforan and 100 mg/l kanamycin. These shoots exhibited GUS activity and Southern analysis showed a single copy insertion into the genome. When the transgenic plants were transferred to soil, they displayed the phenotype specific to the transgenic plants by A. rhizogenes such as dwarfness, delay of flowering, and wrinkled leaves and petals.

  14. AGROBACTERIUM-MEDIATED GENETIC TRANSFORMATION OF SORGHUM USING TISSUE CULTURE-BASED AND POLLEN-MEDIATED APPROACHES

    OpenAIRE

    Elkonin L.A.; O.N. Nosova; J.V. Italianskaya

    2012-01-01

    Genetic transformation is a powerful tool for genetic improvement of arable crops. Genetic engineering approaches are especially important for modification of starch and protein contents, vitamin and micronutrient concentration, improvement of nutritive value of protein fractions, and increase tolerance to environmental stresses. Application of transgenic technologies for genetic improvement of sorghum, a highly productive heat tolerant and drought resistant crop, is extremely important since...

  15. Genetic transformation of the yeast Dekkera/Brettanomyces bruxellensis with non-homologous DNA.

    Science.gov (United States)

    Miklenić, Marina; Štafa, Anamarija; Bajić, Ana; Žunar, Bojan; Lisnić, Berislav; Svetec, Ivan-Krešimir

    2013-05-01

    Yeast Dekkera/Brettanomyces bruxellensis is probably the most common contaminant in wineries and ethanol production processes. The considerable economic losses caused by this yeast, but also its ability to produce and tolerate high ethanol concentrations, make it an attractive subject for research with potential for industrial applications. Unfortunately, efforts to understand the biology of D. bruxellensis and facilitate its broader use in industry are hampered by the lack of adequate procedures for delivery of exogenous DNA into this organism. Here we describe the development of transformation protocols (spheroplast transformation, LiAc/PEG method, and electroporation) and report the first genetic transformation of yeast D. bruxellensis. A linear heterologous DNA fragment carrying the kanMX4 sequence was used for transformation, which allowed transformants to be selected on plates containing geneticin. We found the spheroplast transformation method using 1M sorbitol as osmotic stabilizer to be inappropriate because sorbitol strikingly decreases the plating efficiency of both D. bruxellensis spheroplast and intact cells. However, we managed to modify the LiAc/ PEG transformation method and electroporation to accommodate D. bruxellensis transformation, achieving efficiencies of 0.6-16 and 10-20 transformants/microg DNA, respectively. The stability of the transformants ranged from 93.6% to 100%. All putative transformants were analyzed by Southern blot using the kanMX4 sequence as a hybridization probe, which confirmed that the transforming DNA fragment had integrated into the genome. The results of the molecular analysis were consistent with the expected illegitimate integration of a heterologous transforming fragment.

  16. High-Voltage Electroporation of Bacteria: Genetic Transformation of Campylobacter jejuni with Plasmid DNA

    Science.gov (United States)

    Miller, Jeff F.; Dower, William J.; Tompkins, Lucy S.

    1988-02-01

    Electroporation permits the uptake of DNA by mammalian cells and plant protoplasts because it induces transient permeability of the cell membrane. We investigated the utility of high-voltage electroporation as a method for genetic transformation of intact bacterial cells by using the enteric pathogen Campylobacter jejuni as a model system. This report demonstrates that the application of high-voltage discharges to bacterial cells permits genetic transformation. Our method involves exposure of a Campylobacter cell suspension to a high-voltage exponential decay discharge (5-13 kV/cm) for a brief period of time (resistance-capacitance time constant = 2.4-26 msec) in the presence of plasmid DNA. Electrical transformation of C. jejuni results in frequencies as high as 1.2 × 106 transformants per μ g of DNA. We have investigated the effects of pulse amplitude and duration, cell growth conditions, divalent cations, and DNA concentration on the efficiency of transformation. Transformants of C. jejuni obtained by electroporation contained structurally intact plasmid molecules. In addition, evidence is presented that indicates that C. jejuni possesses DNA restriction and modification systems. The use of electroporation as a method for transforming other bacterial species and guidelines for its implementation are also discussed.

  17. STUDY REGARDING EFFICIENCY OF INDUCED GENETIC TRANSFORMATION IN BACILLUS LICHENIFORMIS WITH PLASMID DNA

    Directory of Open Access Journals (Sweden)

    VINTILĂ T.

    2007-01-01

    Full Text Available A strain of Bacillus licheniformis was subject to genetic transformation with plasmidvectors (pLC1 and pNC61, using electroporation technique, protoplasttransformation and bivalent cations (CaCl2 mediated transformation. In the case oftransformation by electroporation of Bacillus licheniformis B40, the highest numberof transformed colonies (3 were obtained only after a 1,79 KV electric shock, for 2,2milliseconds. Using this transformation technique we have obtained six kanamycinresistant transformants. The frequency of Bacillus licheniformis B40 protoplaststransformation using pLC1 and pNC61 plasmid vectors is approximately 10% (TF =10%. As a result of pLC1 plasmid integration in Bacillus licheniformis protoplasts,six kanamycin resistant transformants were obtained. The pNC61 plasmid, whichconfers trimethoprim resistance, does not integrate in receiver cells by protoplasttransformation. The direct genetic transformation in the presence of bivalent cations(CaCl2, mediated by pLC1 and pNC61 plasmid vectors, produce a lowtransformation frequency. Using this technique, we have obtained three trimethoprimresistant colonies and four kanamycin resistant colonies. The chemical way oftransformation is the only technique, which realizes the integration of pNC61 in B.licheniformis B40 cells.

  18. Genetic transformation of marine cyanobacterium Synechococcus sp. CC9311 (Cyanophyceae) by electroporation

    Science.gov (United States)

    Chen, Huaxin; Lin, Hanzhi; Jiang, Peng; Li, Fuchao; Qin, Song

    2013-03-01

    Synechococcus sp. CC9311 is a marine cyanobacterium characterized by type IV chromatic acclimation (CA). A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA. The results show that Synechococcus sp. CC9311 cells were sensitive to four commonly used antibiotics: ampicillin, kanamycin, spectinomycin, and chloramphenicol. An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV, using a kanamycin resistance gene as selectable marker, was constructed by recombinant polymerase chain reaction. The plasmid was then transformed into Synechococcus sp. CC9311 via electroporation. High transformation efficiency was achieved at a field strength of 2 kV/cm. DNA analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of kanamycin. In addition, the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.

  19. Genetic transformation of marine cyanobacterium Synechococcus sp.CC9311 (Cyanophyceae) by electroporation

    Institute of Scientific and Technical Information of China (English)

    CHEN Huaxin; LIN Hanzhi; JIANG Peng; LI Fuchao; QIN Song

    2013-01-01

    Synechococcus sp.CC9311 is a marine cyanobacterium characterized by type V chromatic acclimation (CA).A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA.The results show that Synechococcus sp.CC9311 cells were sensitive to four commonly used antibiotics:ampicillin,kanamycin,spectinomycin,and chloramphenicol.An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV,using a kanamycin resistance gene as selectable marker,was constructed by recombinant polymerase chain reaction.The plasmid was then transformed into Synechococcus sp.CC9311 via electroporation.High transformation efficiency was achieved at a field strength of 2 kV/cm.DNA analysis showed that rpe V was fully disrupted following challenge of the transformants with a high concentration of kanamycin.In addition,the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.

  20. Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum

    Institute of Scientific and Technical Information of China (English)

    Ji-ye WANG; Hong-ye LI

    2008-01-01

    Agrobacterium tumefaciens-mediated transformation (ATMT) system was assessed for conducting insertional mutagenesis in Penicillium digitatum, a major fungal pathogen infecting post-harvest citrus fruits. A transformation efficiency of up to 60 transformants per 106 conidia was achieved by this system. The integration of the hph gene into the fungal genome was verified by polymerase chain reaction (PCR) amplification and sequencing. These transformants tested were also shown to be mitotically stable. Southern blot analysis of 14 randomly selected transformants showed that the hph gene was randomly integrated as single copy into the fungal genome of P. digitatum. Thus, we conclude that ATMT of P. digitatum could be used as an alternatively practical genetic tool for conducting insertional mutagenesis in P. digitatum to study functional genomics.

  1. Role of molecular genetics in transforming diagnosis of diabetes mellitus.

    Science.gov (United States)

    Molven, Anders; Njølstad, Pål R

    2011-04-01

    Most common diseases also run in families as rare, monogenic forms. Diabetes is no exception. Mutations in approximately 20 different genes are now known to cause monogenic diabetes, a disease group that can be subclassified into maturity-onset diabetes of the young, neonatal diabetes and mitochondrial diabetes. In some families, additional features, such as urogenital malformations, exocrine pancreatic dysfunction and neurological abnormalities, are present and may aid the diagnostic classification. The finding of a mutation in monogenic diabetes may have implications for the prediction of prognosis and choice of treatment. Mutations in the GCK gene cause a mild form of diabetes, which seldom needs insulin and has a low risk for complications. By contrast, HNF1A mutations lead to a diabetes form that in severity, treatment and complication risk resembles Type 1 diabetes, although these patients may experience a good effect of sulfonylurea treatment. The majority of neonatal diabetes cases are caused by mutations in the K(ATP) channel genes ABCC8 and KCNJ11, and sulfonylurea therapy is then usually superior to insulin. Diseases with a considerable genetic component may now be explored by genome-wide approaches using next-generation DNA sequencing technology. We expect that within a few years important breakthroughs will be made in mapping cases of diabetes with a suspected, but still unsolved monogenic basis.

  2. Production of justicidin B, a cytotoxic arylnaphthalene lignan from genetically transformed root cultures of Linum leonii

    NARCIS (Netherlands)

    Vasilev, Nikolay; Elfahmi, [No Value; Bos, Rein; Kayser, Oliver; Momekov, Georgi; Konstantinov, Spiro; Ionkova, Iliana

    2006-01-01

    Callus and hairy root cultures of Linum leonii were established. The genetic transformation in hairy roots was proven by PCR analysis, which showed integration of rol A and rol C genes into the plant genome. Calli and hairy roots accumulate the arylnaphthalene lignan justicidin B as a major constitu

  3. Agrobacterium tumefaciens-mediated genetic transformation of haptophytes (Isochrysis species).

    Science.gov (United States)

    Prasad, Binod; Vadakedath, Nithya; Jeong, Hyun-Jeong; General, Thiyam; Cho, Man-Gi; Lein, Wolfgang

    2014-10-01

    Isochrysis galbana and Isochrysis sp. are economically important microalgae from the division of haptophytes. Here, we report Agrobacterium-mediated stable DNA transfer into their nuclear genomes. Initial studies were performed to standardize co-cultivation media and determine the sensitivity of the microalgae to selective agents. Up to 1 mg/ml of the antibiotic hygromycin did not inhibit growth, whereas both the haptophytes bleached in artificial seawater (ASW) medium containing micromolar concentrations of the herbicide norflurazon. Co-cultivation of Isochrysis sp. and I. galbana with Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pCAMBIA 1380-pds-L504R yielded norflurazon-resistant (NR) colonies visible on selective plates after 20-30 days. pCAMBIA 1380-pds-L540R was constructed by cloning a mutated genomic phytoene desaturase (pds) gene from Haematococcus pluvialis as a selectable marker gene into the binary vector system pCAMBIA 1380. Co-cultivation of Isochrysis sp. with A. tumefaciens in ASW medium containing 200 μM of acetosyringone for 72 h produced the highest number of NR cells. For I. galbana, 100 μM of acetosyringone, ASW medium, and 48 h co-cultivation period appeared to be optimum co-cultivation parameters. The NR colonies kept their resistance phenotype for at least 24 months, even in the absence of selective pressure. The transfer of the pds gene in NR cells was shown by PCR amplification of the T-DNA sequences from the genomic DNA of NR cells and Southern blot analysis using T-DNA sequences as probes. The genetic manipulation described here will allow metabolic engineering and a better understanding of several biochemical pathways in the future.

  4. Toward genetic transformation of mitochondria in mammalian cells using a recoded drug-resistant selection marker

    Institute of Scientific and Technical Information of China (English)

    Young Geol Yoon; Michael Duane Koob

    2011-01-01

    Due to technical difficulties, the genetic transformation of mitochondria in mammalian cells is still a challenge. In this report, we described our attempts to transform mammalian mitochondria with an engineered mitochondrial genome based on selection using a drug resistance gene. Because the standard drug-resistant neomycin phosphotransferase confers resistance to high concentrations of G418 when targeted to the mitochondria, we generated a recoded neomycin resistance gene that uses the mammalian mitochondrial genetic code to direct the synthesis of this protein in the mitochondria, but not in the nucleus (mitochondrial version). We also generated a universal version of the recoded neomycin resistance gene that allows synthesis of the drug-resistant proteins both in the mitochondria and nucleus. When we transfected these recoded neomycin resistance genes that were incorporated into the mouse mitochondrial genome clones into mouse tissue culture cells by electroporation, no DNA constructs were delivered into the mitochondria. We found that the universal version of the recoded neomycin resistance gene was expressed in the nucleus and thus conferred drug resistance to G418 selection, while the synthetic mitochondrial version of the gene produced no background drug-resistant cells from nuclear transformation. These recoded synthetic drug-resistant genes could be a useful tool for selecting mitochondrial genetic transformants as a precise technology for mitochondrial transformation is developed.

  5. [Development of genetic transformation system of Valsa mali of apple mediated by PEG].

    Science.gov (United States)

    Gao, Jing; Li, Yanbo; Ke, Xiwang; Kang, Zhensheng; Huang, Lili

    2011-09-01

    The genetic transformation of Valsa mali var. mali was developed by PEG-mediated protoplasts transformation. It was transformed by PEG-induced fusion of protoplasts. The plasmid pBIG2RHPH2-GFP-GUS carrying hph gene was used and Valsa mali var. mali 03-8 isolate was used as the host strain. At 50 mg/mL driselase + 10 mg/mL lysing enzymes concentration, the mycelium of Valsa mali var. mali cultured in YEPD medium for 48 h was hydrolyzed in 10 mL enzymes liquid /0.5 g wet mycelium for 2 h. The protoplast yield was 4 x 10(7) CFU/mg. The transformation efficiency was 44 per g DNA. Analysis of the transformants by PCR and Southern blotting showed that the selectable marker gene hph was integrated effectively into the genome of Valsa mali var. mali. After 5 subculturing on PDA, 87.5% transformants could grow. This stability test of transformants suggested that the foreign gene hph was stable in heredity. This transformation system is a valuable and important tool for the further study of the pathogenic gene of Valsa mali.

  6. A genetic-algorithm-based method to find the unitary transformations for any de- sired quantum computation and application to a one-bit oracle decision problem

    OpenAIRE

    Bang, Jeongho; Yoo, Seokwon

    2014-01-01

    We propose a genetic-algorithm-based method to find the unitary transformations for any desired quantum computation. We formulate a simple genetic algorithm by introducing the "genetic parameter vector" of the unitary transformations to be found. In the genetic algorithm process, all components of the genetic parameter vectors are supposed to evolve to the solution parameters of the unitary transformations. We apply our method to find the optimal unitary transformations and to generalize the ...

  7. Improvement of tissue culture, genetic transformation, and applications of biotechnology to Brassica.

    Science.gov (United States)

    Ravanfar, Seyed Ali; Orbovic, Vladimir; Moradpour, Mahdi; Abdul Aziz, Maheran; Karan, Ratna; Wallace, Simon; Parajuli, Saroj

    2017-04-01

    Development of in vitro plant regeneration method from Brassica explants via organogenesis and somatic embryogenesis is influenced by many factors such as culture environment, culture medium composition, explant sources, and genotypes which are reviewed in this study. An efficient in vitro regeneration system to allow genetic transformation of Brassica is a crucial tool for improving its economical value. Methods to optimize transformation protocols for the efficient introduction of desirable traits, and a comparative analysis of these methods are also reviewed. Hence, binary vectors, selectable marker genes, minimum inhibitory concentration of selection agents, reporter marker genes, preculture media, Agrobacterium concentration and regeneration ability of putative transformants for improvement of Agrobacterium-mediated transformation of Brassica are discussed.

  8. Degradation of endocrine disrupting chemicals by genetic transformants with two lignin degrading enzymes in Phlebia tremellosa.

    Science.gov (United States)

    Kum, Hyunwoo; Lee, Sungsuk; Ryu, Sunhwa; Choi, Hyoung T

    2011-10-01

    A white rot fungus Phlebia tremellosa produced lignin degrading enzymes, which showed degrading activity against various recalcitrant compounds. However, manganese peroxidase (MnP) activity, one of lignin degrading enzymes, was very low in this fungus under various culture conditions. An expression vector that carried both the laccase and MnP genes was constructed using laccase genomic DNA of P. tremellosa and MnP cDNA from Polyporus brumalis. P. tremellosa was genetically transformed using the expression vector to obtain fungal transformants showing increased laccase and MnP activity. Many transformants showed highly increased laccase and MnP activity at the same time in liquid medium, and three of them were used to degrade endocrine disrupting chemicals. The transformant not only degraded bisphenol A and nonylphenol more rapidly but also removed the estrogenic activities of the chemicals faster than the wild type strain.

  9. Abdomen disease diagnosis in CT images using flexiscale curvelet transform and improved genetic algorithm.

    Science.gov (United States)

    Sethi, Gaurav; Saini, B S

    2015-12-01

    This paper presents an abdomen disease diagnostic system based on the flexi-scale curvelet transform, which uses different optimal scales for extracting features from computed tomography (CT) images. To optimize the scale of the flexi-scale curvelet transform, we propose an improved genetic algorithm. The conventional genetic algorithm assumes that fit parents will likely produce the healthiest offspring that leads to the least fit parents accumulating at the bottom of the population, reducing the fitness of subsequent populations and delaying the optimal solution search. In our improved genetic algorithm, combining the chromosomes of a low-fitness and a high-fitness individual increases the probability of producing high-fitness offspring. Thereby, all of the least fit parent chromosomes are combined with high fit parent to produce offspring for the next population. In this way, the leftover weak chromosomes cannot damage the fitness of subsequent populations. To further facilitate the search for the optimal solution, our improved genetic algorithm adopts modified elitism. The proposed method was applied to 120 CT abdominal images; 30 images each of normal subjects, cysts, tumors and stones. The features extracted by the flexi-scale curvelet transform were more discriminative than conventional methods, demonstrating the potential of our method as a diagnostic tool for abdomen diseases.

  10. Novel insights in genetic transformation of the probiotic yeast Saccharomyces boulardii

    Science.gov (United States)

    Douradinha, Bruno; Reis, Viviane CB; Rogers, Matthew B; Torres, Fernando AG; Evans, Jared D; Marques Jr, Ernesto TA

    2014-01-01

    Saccharomyces boulardii (S. boulardii) is a probiotic yeast related to Saccharomyces cerevisiae (S. cerevisiae) but with distinct genetic, taxonomic and metabolic properties. S. cerevisiae has been used extensively in biotechnological applications. Currently, many strains are available, and multiple genetic tools have been developed, which allow the expression of several exogenous proteins of interest with applications in the fields of medicine, biofuels, the food industry, and scientific research, among others. Although S. boulardii has been widely studied due to its probiotic properties against several gastrointestinal tract disorders, very few studies addressed the use of this yeast as a vector for expression of foreign genes of interest with biotechnological applications. Here we show that, despite the similarity of the two yeasts, not all genetic tools used in S. cerevisiae can be applied in S. boulardii. While transformation of the latter could be obtained using a commercial kit developed for the former, consequent screening of successful transformants had to be optimized. We also show that several genes frequently used in genetic manipulation of S. cerevisiae (e.g., promoters and resistance markers) are present in S. boulardii. Sequencing revealed a high rate of homology (>96%) between the orthologs of the two yeasts. However, we also observed some of them are not eligible to be targeted for transformation of S. boulardii. This work has important applications toward the potential of this probiotic yeast as an expression system for genes of interest. PMID:24013355

  11. Combined genetic transformation and nutritional assay for identification of Moraxella nonliquefaciens.

    Science.gov (United States)

    Juni, E; Heym, G A; Maurer, M J; Miller, M L

    1987-09-01

    A combined genetic transformation and nutritional assay is described that permits definitive identification of clinically isolated strains of Moraxella nonliquefaciens. Crude DNA preparations of strains of various Moraxella species were used to transform nutritional mutants of a stably competent strain of M. nonliquefaciens for ability to grow on a defined medium (Mn-B). DNA samples from 24 independently isolated strains of M. nonliquefaciens all resulted in massive (4+) transformation of each of two mutant assay strains. DNA samples from strains of M. bovis and M. lacunata frequently gave weak (1+) transformation of one of the mutant assay strains (Mn64) but almost always failed to transform another assay strain (Mn136). DNA samples from eight other Moraxella species failed completely to transform either of the mutant assay strains. When streaked on the defined medium used for the transformation assay (Mn-B), 23 of the 24 strains of M. nonliquefaciens grew well, but all strains of M. bovis and M. lacunata failed to grow on this medium.

  12. Embryogenic callus proliferation and regeneration conditions for genetic transformation of diverse sugarcane cultivars.

    Science.gov (United States)

    Basnayake, Shiromani W V; Moyle, Richard; Birch, Robert G

    2011-03-01

    Amenability to tissue culture stages required for gene transfer, selection and plant regeneration are the main determinants of genetic transformation efficiency via particle bombardment into sugarcane. The technique is moving from the experimental phase, where it is sufficient to work in a few amenable genotypes, to practical application in a diverse and changing set of elite cultivars. Therefore, we investigated the response to callus initiation, proliferation, regeneration and selection steps required for microprojectile-mediated transformation, in a diverse set of Australian sugarcane cultivars. 12 of 16 tested cultivars were sufficiently amenable to existing routine tissue-culture conditions for practical genetic transformation. Three cultivars required adjustments to 2,4-D levels during callus proliferation, geneticin concentration during selection, and/or light intensity during regeneration. One cultivar gave an extreme necrotic response in leaf spindle explants and produced no callus tissue under the tested culture conditions. It was helpful to obtain spindle explants for tissue culture from plants with good water supply for growth, especially for genotypes that were harder to culture. It was generally possible to obtain several independent transgenic plants per bombardment, with time in callus culture limited to 11-15 weeks. A caution with this efficient transformation system is that separate shoots arose from different primary transformed cells in more than half of tested calli after selection for geneticin resistance. The results across this diverse cultivar set are likely to be a useful guide to key variables for rapid optimisation of tissue culture conditions for efficient genetic transformation of other sugarcane cultivars.

  13. Genome-wide profiling of genetic variation in Agrobacterium-transformed rice plants*#

    Science.gov (United States)

    Li, Wen-xu; Wu, San-ling; Liu, Yan-hua; Jin, Gu-lei; Zhao, Hai-jun; Fan, Long-jiang; Shu, Qing-yao

    2016-01-01

    Agrobacterium-mediated transformation has been widely used in producing transgenic plants, and was recently used to generate “transgene-clean” targeted genomic modifications coupled with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas9) system. Although tremendous variation in morphological and agronomic traits, such as plant height, seed fertility, and grain size, was observed in transgenic plants, the underlying mechanisms are not yet well understood, and the types and frequency of genetic variation in transformed plants have not been fully disclosed. To reveal the genome-wide variation in transformed plants, we sequenced the genomes of five independent T0 rice plants using next-generation sequencing (NGS) techniques. Bioinformatics analyses followed by experimental validation revealed the following: (1) in addition to transfer-DNA (T-DNA) insertions, three transformed plants carried heritable plasmid backbone DNA of variable sizes (855–5216 bp) and in different configurations with the T-DNA insertions (linked or apart); (2) each transgenic plant contained an estimated 338–1774 independent genetic variations (single nucleotide variations (SNVs) or small insertion/deletions); and (3) 2–6 new Tos17 insertions were detected in each transformed plant, but no other transposable elements or bacterial genomic DNA. PMID:27921404

  14. Genetic transformation with the gfp gene of Colletotrichum gloeosporioides isolates from coffee with blister spot.

    Science.gov (United States)

    Armesto, Cecilia; Maia, Fernanda Gonçalves Martins; de Abreu, Mário Sobral; Figueira, Antonia Dos Reis; da Silva, Bruno Marques; Monteiro, Fernando Pereira

    2012-07-01

    Blister spot (Colletotrichum gloeosporioides) is now widespread in most coffee producing states of Brazil, becoming a limiting factor for production. The lack of data relating to the reproduction of typical symptoms (light green, oily patches) leaves a gap within the pathosystem, forcing the search for new methodologies for monitoring the disease. Monitoring of genetically modified organisms has proven to be an effective tool in understanding the host × pathogen interactions. Thus, the present study was carried out to evaluate the effectiveness of two systems of genetic transformation in obtaining mutants using the gfp reporter gene. Using the two transformation systems (PEG and electroporation) revealed the efficiency of both, confirmed by fluorescence microscopy and resistance to the antibiotic hygromycin-B, when incorporated into the culture medium. The fungus maintained its cultural and morphological characteristics when compared to wild strains. When inoculated on coffee seedlings, it was found that the pathogenicity of the processed isolates had not changed.

  15. Genetic transformation with the gfp gene of Colletotrichum gloeosporioides isolates from coffee with blister spot

    Directory of Open Access Journals (Sweden)

    Cecilia Armesto

    2012-09-01

    Full Text Available Blister spot (Colletotrichum gloeosporioides is now widespread in most coffee producing states of Brazil, becoming a limiting factor for production. The lack of data relating to the reproduction of typical symptoms (light green, oily patches leaves a gap within the pathosystem, forcing the search for new methodologies for monitoring the disease. Monitoring of genetically modified organisms has proven to be an effective tool in understanding the host x pathogen interactions. Thus, the present study was carried out to evaluate the effectiveness of two systems of genetic transformation in obtaining mutants using the gfp reporter gene. Using the two transformation systems (PEG and electroporation revealed the efficiency of both, confirmed by fluorescence microscopy and resistance to the antibiotic hygromycin-B, when incorporated into the culture medium. The fungus maintained its cultural and morphological characteristics when compared to wild strains. When inoculated on coffee seedlings, it was found that the pathogenicity of the processed isolates had not changed.

  16. Inrush Current Simulation of Power Transformer using Machine Parameters Estimated by Design Procedure of Winding Structure and Genetic Algorithm

    Science.gov (United States)

    Tokunaga, Yoshitaka

    This paper presents estimation techniques of machine parameters for power transformer using design procedure of transformer and genetic algorithm with real coding. Especially, it is very difficult to obtain machine parameters for transformers in customers' facilities. Using estimation techniques, machine parameters could be calculated from the only nameplate data of these transformers. Subsequently, EMTP-ATP simulation of the inrush current was carried out using machine parameters estimated by techniques developed in this study and simulation results were reproduced measured waveforms.

  17. The Technique of Genetic Transformation Mediated by keV Ion Beam

    Institute of Scientific and Technical Information of China (English)

    卞坡; 余增亮

    2005-01-01

    The application of keV ion beam in life science started in China several decades ago. In 1986, researchers initially studied the mutagenic effect of ion beam, and successfully applied it to plant breeding. Nowadays, ion beam implantation technique has been extensively applied to many biological fields. This paper mainly introduces one of its important applications: genetic transformation mediated by keV ion beam.

  18. An improved Agrobacterium-mediated transformation system for the functional genetic analysis of Penicillium marneffei.

    Science.gov (United States)

    Kummasook, Aksarakorn; Cooper, Chester R; Vanittanakom, Nongnuch

    2010-12-01

    We have developed an improved Agrobacterium-mediated transformation (AMT) system for the functional genetic analysis of Penicillium marneffei, a thermally dimorphic, human pathogenic fungus. Our AMT protocol included the use of conidia or pre-germinated conidia of P. marneffei as the host recipient for T-DNA from Agrobacterium tumefaciens and co-cultivation at 28°C for 36 hours. Bleomycin-resistant transformants were selected as yeast-like colonies following incubation at 37°C. The efficiency of transformation was approximately 123 ± 3.27 and 239 ± 13.12 transformants per plate when using 5 × 10(4) conidia and pre-germinated conidia as starting materials, respectively. Southern blot analysis demonstrated that 95% of transformants contained single copies of T-DNA. Inverse PCR was employed for identifying flanking sequences at the T-DNA insertion sites. Analysis of these sequences indicated that integration occurred as random recombination events. Among the mutants isolated were previously described stuA and gasC defective strains. These AMT-derived mutants possessed single T-DNA integrations within their particular coding sequences. In addition, other morphological and pigmentation mutants possessing a variety of gene-specific defects were isolated, including two mutants having T-DNA integrations within putative promoter regions. One of the latter integration events was accompanied by the deletion of the entire corresponding gene. Collectively, these results indicated that AMT could be used for large-scale, functional genetic analyses in P. marneffei. Such analyses can potentially facilitate the identification of those genetic elements related to morphogenesis, as well as pathogenesis in this medically important fungus.

  19. Quasi-conformal mapping with genetic algorithms applied to coordinate transformations

    Science.gov (United States)

    González-Matesanz, F. J.; Malpica, J. A.

    2006-11-01

    In this paper, piecewise conformal mapping for the transformation of geodetic coordinates is studied. An algorithm, which is an improved version of a previous algorithm published by Lippus [2004a. On some properties of piecewise conformal mappings. Eesti NSV Teaduste Akademmia Toimetised Füüsika-Matemaakika 53, 92-98; 2004b. Transformation of coordinates using piecewise conformal mapping. Journal of Geodesy 78 (1-2), 40] is presented; the improvement comes from using a genetic algorithm to partition the complex plane into convex polygons, whereas the original one did so manually. As a case study, the method is applied to the transformation of the Spanish datum ED50 and ETRS89, and both its advantages and disadvantages are discussed herein.

  20. SUFISTIC AND TRANSFORMATIVE PEDAGOGIC VALUES IN SYAIKH SITI JENAR NOVEL BY AGUS SUNYOTO GENETIC STRUCTURALISM

    Directory of Open Access Journals (Sweden)

    Sugeng Riadi

    2015-06-01

    Full Text Available This research aims to know and understand more deeply about mystical values and transformative pedagogic values in the Syaikh Siti Jenar novel written by Agus Sunyoto. The focus of this study was the intrinsic and extrinsic structure of meaning in the novel. The method applied in this study is  content analysis  using genetic structure by Lucien Goldmann. The results of the study show a number of sufistic values and transformative pedagogic values. Sufistic values include: repentance, ascetic, indigent, patience, gratitude, pleasure, and trust. While the transformative pedagogic values include altruism, egalitarian, pluralism, and eclectic. The findings of the author's world view includes figures deconstruction of the teachings of Syaikh Siti Jenar, cultural missionary endeavor, the concept of superhuman, mystical literature, and literary history.

  1. Establishment of an Indirect Genetic Transformation Method for Arabidopsis thaliana ecotype Bangladesh

    Directory of Open Access Journals (Sweden)

    Bulbul AHMED

    2011-11-01

    Full Text Available Arabidopsis thaliana is a small flowering plant belonging to the Brassicaceae family, which is adopted as a model plant for genetic research. Agrobacterium tumifaciensmediated transformation method for A. thaliana ecotype Bangladesh was established. Leaf discs of A. thaliana were incubated with A. tumefaciens strain LBA4404 containing chimeric nos. nptII. nos and intron-GUS genes. Following inoculation and co-cultivation, leaf discs were cultured on selection medium containing 50 mg/l kanamycin + 50 mg/l cefotaxime + 1.5 mg/l NAA and kanamycin resistant shoots were induced from the leaf discs after two weeks. Shoot regeneration was achieved after transferring the tissues onto fresh medium of the same combination. Finally, the shoots were rooted on MS medium containing 50 mg/l kanamycin. Incorporation and expression of the transgenes were confirmed by PCR analysis. Using this protocol, transgenic A. thaliana plants can be obtained and indicates that genomic transformation in higher plants is possible through insertion of desired gene. Although Agrobacterium mediated genetic transformation is established for A. thaliana, this study was the conducted to transform A. thaliana ecotype Bangladesh.

  2. Agrobacterium-mediated genetic transformation of pineapple (Ananas comosus L., Merr.).

    Science.gov (United States)

    Mhatre, Minal

    2013-01-01

    Pineapple (Ananas comosus L., Merr.) is a commercially important crop, grown in the tropical and subtropical regions. However, the crop is faced with postharvest damage and poor varietal and nutritional improvement. Being a vegetatively propagated crop, conventional breeding programs take longer time for genetic improvement, which may not necessarily successfully develop an improved cultivar. Hence, the genetic modification of pineapple is an alternative handy approach to improve pineapple. We have established an Agrobacterium-mediated transformation system using leaf bases from in vitro-grown pineapple plants. Being a monocot, acetosyringone is added to the culture medium for overnight growth of Agrobacterium and transformation to transfer a gene of interest MSI99 soybean ferritin. Leaf bases isolated from in vitro shoot cultures are treated with Agrobacterium suspension at two dilutions, 10× and 20×, for 30 min. Explants are subsequently blot dried and cultured on gelrite solidified hormone-free Pin1 medium for 2 days (cocultivation). Periodic transfer is first done to the regeneration medium (Pin1) containing cefotaxime for the suppression of Agrobacterium growth. The transformants are selected by culturing on Pin1 medium containing cefotaxime and kanamycin. Multiple shoots, regenerated in leaf bases, are further multiplied and individually rooted in the liquid RM medium amended with antibiotics to recover plants. Putative transformants are analyzed for transgene integration and expression using standard molecular biological methods of PCR, RT-PCR, and genomic Southern.

  3. Genetic transformation of Nannochloropsis oculata with a bacterial phleomycin resistance gene as dominant selective marker

    Science.gov (United States)

    Ma, Xiaolei; Pan, Kehou; Zhang, Lin; Zhu, Baohua; Yang, Guanpin; Zhang, Xiangyang

    2016-04-01

    The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×107 cells mL-1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL-1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.

  4. Analysis of a genetically structured variance heterogeneity model using the Box-Cox transformation

    DEFF Research Database (Denmark)

    Yang, Ye; Christensen, Ole Fredslund; Sorensen, Daniel

    2011-01-01

    of the marginal distribution of the data. To investigate how the scale of measurement affects inferences, the genetically structured heterogeneous variance model is extended to accommodate the family of Box–Cox transformations. Litter size data in rabbits and pigs that had previously been analysed...... in the untransformed scale were reanalysed in a scale equal to the mode of the marginal posterior distribution of the Box–Cox parameter. In the rabbit data, the statistical evidence for a genetic component at the level of the environmental variance is considerably weaker than that resulting from an analysis...... in the original metric. In the pig data, the statistical evidence is stronger, but the coefficient of correlation between additive genetic effects affecting mean and variance changes sign, compared to the results in the untransformed scale. The study confirms that inferences on variances can be strongly affected...

  5. Optimization of Agrobacterium mediated genetic transformation of cotyledonary node explants of Vigna radiata.

    Science.gov (United States)

    Yadav, Sushil Kumar; Katikala, Sweety; Yellisetty, Varalaxmi; Kannepalle, Annapurna; Narayana, Jyothi Lakshmi; Maddi, Vanaja; Mandapaka, Maheswari; Shanker, Arun Kumar; Bandi, Venkateswarlu; Bharadwaja, Kirti Pulugurtha

    2012-12-01

    A reproducible and highly efficient protocol for genetic transformation mediated by Agrobacterium has been established for greengram (Vigna radiata L. Wilczek). Double cotyledonary node (DCN) explants were inoculated with Agrobacterium tumefaciens strain LBA 4404 harboring a binary vector pCAMBIA 2301 containing neomycin phosphotransferase (npt II) gene as selectable marker, β-glucuronidase (GUS) as a reporter (uidA) gene and annexin 1 bj gene. Important parameters like optical density of Agrobacterium culture, culture quantity, infection medium, infection and co-cultivation time and acetosyringone concentration were standardized to optimize the transformation frequency. Kanamycin at a concentration of 100 mg/l was used to select transformed cells. Transient and stable GUS expressions were studied in transformed explants and regenerated putative plants, respectively. Transformed shoot were produced on regeneration medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime and rooted on ½ MS medium. Transient and constitutive GUS expression was observed in DCN explants and different tissues of T(0) and T(1) plants. Rooted T(0) and T(1) shoots confirming Polymerase Chain Reaction (PCR) positive for npt II and annexin 1bj genes were taken to maturity to collect the seeds. Integration of annexin gene into the greengram genome was confirmed by Southern blotting.

  6. A High-Throughput Regeneration and Transformation Platform for Production of Genetically Modified Banana.

    Science.gov (United States)

    Tripathi, Jaindra N; Oduor, Richard O; Tripathi, Leena

    2015-01-01

    Banana (Musa spp.) is an important staple food as well as cash crop in tropical and subtropical countries. Various bacterial, fungal, and viral diseases and pests such as nematodes are major constraints in its production and are currently destabilizing the banana production in sub-Saharan Africa. Genetic engineering is a complementary option used for incorporating useful traits in banana to bypass the long generation time, polyploidy, and sterility of most of the cultivated varieties. A robust transformation protocol for farmer preferred varieties is crucial for banana genomics and improvement. A robust and reproducible system for genetic transformation of banana using embryogenic cell suspensions (ECS) has been developed in this study. Two different types of explants (immature male flowers and multiple buds) were tested for their ability to develop ECS in several varieties of banana locally grown in Africa. ECS of banana varieties "Cavendish Williams" and "Gros Michel" were developed using multiple buds, whereas ECS of "Sukali Ndiizi" was developed using immature male flowers. Regeneration efficiency of ECS was about 20,000-50,000 plantlets per ml of settled cell volume (SCV) depending on variety. ECS of three different varieties were transformed through Agrobacterium-mediated transformation using gusA reporter gene and 20-70 independent transgenic events per ml SCV of ECS were regenerated on selective medium. The presence and integration of gusA gene in transgenic plants was confirmed by PCR, dot blot, and Southern blot analysis and expression by histochemical GUS assays. The robust transformation platform was successfully used to generate hundreds of transgenic lines with disease resistance. Such a platform will facilitate the transfer of technologies to national agricultural research systems (NARS) in Africa.

  7. Meropenem as an Alternative Antibiotic Agent for Suppression of Agrobacterium in Genetic Transformation of Orchid

    Institute of Scientific and Technical Information of China (English)

    CAO Ying; Niimi Yoshiyuki; HU Shang-lian

    2006-01-01

    A case of Meropenem as a novel antibacterial agent to suppress and eliminate Agrobacterium tumefaciens in the Agrobacterium-mediated transformation of orchid protocorm-like bodies (PLBs) has been reported in this article. The in vitro activities of meropenem and four comparator antibacterial agents against three Agrobacterium tumefaciens strains, LBA4404, EHA101, and GV3101, were assessed. In addition, the effect of meropenem on the growth of Dendrobium phalaenopsis PLBs was determined. Compared with other commonly used antibiotics (including ampicillin,carbenicillin, cefotaxime, and cefoperazone), meropenem showed the highest activity in suppressing all tested A.tumefaciens strains (minimum inhibitory concentration [MIC] < 0.5 mg L-1, which is equal to minimum bactericidal concentration [MBC]). Meropenem, at all tested concentrations, except for 10 mg L-1 concentration, had little negative effect on the growth of orchid tissues. The A. tumefaciens strain EHA101 in genetic transformation with vector pIG121Hm in infected PLBs of the orchid was visually undetectable after a two-month subculture in 1/2 MS medium with 50 mg L-1 meropenem and 25 mg L-1 hygromacin. The expression and incorporation of the transgenes were confirmed by GUS histochemical assay and PCR analysis. Meropenem may be an alternative antibiotic for the effective suppression of A. tumefaciens in genetic transformation.

  8. Genetic and environmental information in goat milk Fourier transform infrared spectra.

    Science.gov (United States)

    Dagnachew, B S; Kohler, A; Adnøy, T

    2013-06-01

    Fourier transform infrared (FTIR) spectroscopy is often used in prediction of major milk components in genetic evaluation of dairy animals. Until now genetic variability of goat milk FTIR spectra has only been known indirectly through their contribution to the major milk components. In this study, genetic and environmental components of goat milk FTIR spectra were examined directly. A data set containing 83,858 milk FTIR spectral observations belonging to 29,320 Norwegian dairy goats of 271 herds was used for the study. Principal components analysis was applied on both unprocessed and preprocessed spectral data, and new traits (latent traits) were defined because a multitrait analysis of all spectral variables for variance components could not be done. Eight and 7 latent variables, explaining approximately 99% of the total unprocessed and preprocessed spectral variation, respectively, were kept from the principal components analysis for genetic analysis. Genetic and environmental variance components were estimated for the latent traits using restricted maximum likelihood. Genetic-to-total phenotypic variance ratios (heritabilities) of the latent traits were between 0.011 and 0.285 for the unprocessed spectra and between 0.135 and 0.262 for the preprocessed spectra. The estimated variance components for the latent traits were back transformed to the spectral variables. Heritabilities of these spectral variables ranged from 0.018 to 0.408 and variance ratios of the permanent environmental effects of goats were between 0.002 and 0.184 of the phenotypic spectral variation. High-to-moderate heritabilities were observed in particular in spectral regions related to major milk components (fat, lactose, and protein): between 1,030 and 1,300 cm(-1), 1,500 and 1,600 cm(-1), 1,700 and 1,800 cm(-1), and 2,800 and 3,000 cm(-1). Our results confirmed that a substantial amount of genetic variation exists in goat milk FTIR spectra. Not all spectral variations are of genetic origin

  9. Genetic effects of some platinum co-ordination complexes on E.coli DNA as revealed by transformation studies

    NARCIS (Netherlands)

    Hoekstra, W.P.M.; Daemen, Toos

    1982-01-01

    E. coli chromosomal DNA was treated with various Pt co-ordination compounds and then used as donor DNA in E. coli transformation. Genetic analysis of transformants obtained with Pt-treated DNA showed effects of cis-diamminedichloroplatinum(II) (cis-Pt(II)) and cis-Pt-dimethyl-1,3-diaminopropane Cl4

  10. Assessment of factors affecting Agrobacterium-mediated genetic transformation of the unicellular green alga, Chlorella vulgaris.

    Science.gov (United States)

    Cha, Thye San; Yee, Willy; Aziz, Ahmad

    2012-04-01

    The successful establishment of an Agrobacterium-mediated transformation method and optimisation of six critical parameters known to influence the efficacy of Agrobacterium T-DNA transfer in the unicellular microalga Chlorella vulgaris (UMT-M1) are reported. Agrobacterium tumefaciens strain LBA4404 harbouring the binary vector pCAMBIA1304 containing the gfp:gusA fusion reporter and a hygromycin phosphotransferase (hpt) selectable marker driven by the CaMV35S promoter were used for transformation. Transformation frequency was assessed by monitoring transient β-glucuronidase (GUS) expression 2 days post-infection. It was found that co-cultivation temperature at 24°C, co-cultivation medium at pH 5.5, 3 days of co-cultivation, 150 μM acetosyringone, Agrobacterium density of 1.0 units (OD(600)) and 2 days of pre-culture were optimum variables which produced the highest number of GUS-positive cells (8.8-20.1%) when each of these parameters was optimised individually. Transformation conducted with the combination of all optimal parameters above produced 25.0% of GUS-positive cells, which was almost a threefold increase from 8.9% obtained from un-optimised parameters. Evidence of transformation was further confirmed in 30% of 30 randomly-selected hygromycin B (20 mg L(-1)) resistant colonies by polymerase chain reaction (PCR) using gfp:gusA and hpt-specific primers. The developed transformation method is expected to facilitate the genetic improvement of this commercially-important microalga.

  11. An Image Filter Based on Multiobjective Genetic Algorithm and Shearlet Transformation

    Directory of Open Access Journals (Sweden)

    Zhi-yong Fan

    2013-01-01

    Full Text Available Rician noise pollutes magnetic resonance imaging (MRI data, making data’s postprocessing difficult. In order to remove this noise and avoid loss of details as much as possible, we proposed a filter algorithm using both multiobjective genetic algorithm (MOGA and Shearlet transformation. Firstly, the multiscale wavelet decomposition is applied to the target image. Secondly, the MOGA target function is constructed by evaluation methods, such as signal-to-noise ratio (SNR and mean square error (MSE. Thirdly, MOGA is used with optimal coefficients of Shearlet wavelet threshold value in a different scale and a different orientation. Finally, the noise-free image could be obtained through inverse wavelet transform. At the end of the paper, experimental results show that this proposed algorithm eliminates Rician noise more effectively and yields better peak signal-to-noise ratio (PSNR gains compared with other traditional filters.

  12. Genetic transformation of midgut bacteria from the red imported fire ant (Solenopsis invicta).

    Science.gov (United States)

    Medina, Freder; Li, Haiwen; Vinson, S Bradleigh; Coates, Craig J

    2009-05-01

    In our previous study we isolated 10 bacterial species from fourth-instar larval midguts of the red imported fire ant, Solenopsis invicta. Here we report the genetic transformation and reintroduction of three species (Kluyvera cryocrescens, Serratia marcescens, and isolate 38) into the fire ant host. All three species were transformed with the plasmid vector, pZeoDsRed. High expression levels of DsRed were observed and the plasmid is maintained in these bacteria at 37 degrees C in the absence of antibiotic selection for at least 9 days of subculturing. The transformed bacteria were successfully reintroduced into fire ant larvae and survived in the fire ant gut for at least 7 days. Upon pupal emergence, 7 days after reintroduction, transformed bacteria can still be isolated, however, most were passed out in the meconium. We further demonstrated that the engineered bacteria could be spread within the colony by feeding this meconium to naive larvae with the aid of worker fire ants.

  13. Combinatorial genetic transformation of cereals and the creation of metabolic libraries for the carotenoid pathway.

    Science.gov (United States)

    Farre, Gemma; Naqvi, Shaista; Sanahuja, Georgina; Bai, Chao; Zorrilla-López, Uxue; Rivera, Sol M; Canela, Ramon; Sandman, Gerhard; Twyman, Richard M; Capell, Teresa; Zhu, Changfu; Christou, Paul

    2012-01-01

    Combinatorial nuclear transformation is used to generate populations of transgenic plants containing random selections from a collection of input transgenes. This is a useful approach because it provides the means to test different combinations of genes without the need for separate transformation experiments, allowing the comprehensive analysis of metabolic pathways and other genetic systems requiring the coordinated expression of multiple genes. The principle of combinatorial nuclear transformation is demonstrated in this chapter through protocols developed in our laboratory that allow combinations of genes encoding enzymes in the carotenoid biosynthesis pathway to be introduced into rice and a white-endosperm variety of corn. These allow the accumulation of carotenoids to be screened initially by the colour of the endosperm, which ranges from white through various shades of yellow and orange depending on the types and quantities of carotenoids present. The protocols cover the preparation of DNA-coated metal particles, the transformation of corn and rice plants by particle bombardment, the regeneration of transgenic plants, the extraction of carotenoids from plant tissues, and their analysis by high-performance liquid chromatography.

  14. Genetic transformation of cork oak (Quercus suber L.) for herbicide resistance.

    Science.gov (United States)

    Alvarez, Rubén; Alvarez, José M; Humara, Jaime M; Revilla, Angeles; Ordás, Ricardo J

    2009-09-01

    The bar gene was introduced into the cork oak genome. Cork oak embryogenic masses were transformed using the Agrobacterium strain AGL1 which carried the plasmid pBINUbiBar. This vector harbours the genes, nptII and bar, the latter under control of the maize ubiquitin promoter. The transgenic embryogenic lines were cryopreserved. Varying activities of phosphinothricin acetyl transferase were detected among the lines, which carried 1-4 copies of the insert. Molecular and biochemical assays confirmed the stability and expression of the transgenes 3 months after thawing the cultures. These results demonstrate genetic engineering of herbicide tolerance in Quercus spp.

  15. Handling Arabidopsis plants: growth, preservation of seeds, transformation, and genetic crosses.

    Science.gov (United States)

    Rivero, Luz; Scholl, Randy; Holomuzki, Nicholas; Crist, Deborah; Grotewold, Erich; Brkljacic, Jelena

    2014-01-01

    Growing healthy plants is essential for the advancement of Arabidopsis thaliana (Arabidopsis) research. Over the last 20 years, the Arabidopsis Biological Resource Center (ABRC) has collected and developed a series of best-practice protocols, some of which are presented in this chapter. Arabidopsis can be grown in a variety of locations, growth media, and environmental conditions. Most laboratory accessions and their mutant or transgenic derivatives flower after 4-5 weeks and set seeds after 7-8 weeks, under standard growth conditions (soil, long day, 23 ºC). Some mutant genotypes, natural accessions, and Arabidopsis relatives require strict control of growth conditions best provided by growth rooms, chambers, or incubators. Other lines can be grown in less-controlled greenhouse settings. Although the majority of lines can be grown in soil, certain experimental purposes require utilization of sterile solid or liquid growth media. These include the selection of primary transformants, identification of homozygous lethal individuals in a segregating population, or bulking of a large amount of plant material. The importance of controlling, observing, and recording growth conditions is emphasized and appropriate equipment required to perform monitoring of these conditions is listed. Proper conditions for seed harvesting and preservation, as well as seed quality control, are also described. Plant transformation and genetic crosses, two of the methods that revolutionized Arabidopsis genetics, are introduced as well.

  16. Establishment of a Genetic Transformation System and Its Application in Thermoanaerobacter tengcongensis

    Institute of Scientific and Technical Information of China (English)

    Bo Liu; Chuan Wang; Haihua Yang; Huarong Tan

    2012-01-01

    The whole-genome sequence of Thermoanaerobacter tengcongensis,an anaerobic thermophilic bacterium isolated from the Tengchong hot spring in China,was completed in 2002.However,in vivo studies on the genes of this strain have been hindered in the absence of genetic manipulation system.In order to establish such a system,the plasmid pBOL01 containing the replication origin of the T.tengcongensis chromosome and a kanamycin resistance cassette,in which kanamycin resistance gene expression was controlled by the tte1482 promoter from T.tengcongensis,was constructed and introduced into T.tengcongensis via electroporation.Subsequently,the high transformation efficiency occurred when using freshly cultured T.tengcongensis cells without electroporation treatment,suggesting that T.tengcongensis is naturally competent under appropriate growth stage.A genetic transformation system for this strain was then established based on these important components,and this system was proved to be available for studying physiological characters of T.tengcongensis in vivo by means of hisG gene disruption and complementation.

  17. Transformation

    DEFF Research Database (Denmark)

    Peters, Terri

    2011-01-01

    Artiklen diskuterer ordet "transformation" med udgangspunkt i dels hvorledes ordet bruges i arkitektfaglig terminologi og dels med fokus på ordets potentielle indhold og egnethed i samme teminologi.......Artiklen diskuterer ordet "transformation" med udgangspunkt i dels hvorledes ordet bruges i arkitektfaglig terminologi og dels med fokus på ordets potentielle indhold og egnethed i samme teminologi....

  18. Transformation

    DEFF Research Database (Denmark)

    Peters, Terri

    2011-01-01

    Artiklen diskuterer ordet "transformation" med udgangspunkt i dels hvorledes ordet bruges i arkitektfaglig terminologi og dels med fokus på ordets potentielle indhold og egnethed i samme teminologi.......Artiklen diskuterer ordet "transformation" med udgangspunkt i dels hvorledes ordet bruges i arkitektfaglig terminologi og dels med fokus på ordets potentielle indhold og egnethed i samme teminologi....

  19. In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation

    Directory of Open Access Journals (Sweden)

    Hafiz Mamoon REHMAN

    2012-11-01

    Full Text Available Dalbergia sissoo Roxb. ex DC. (Sissoo is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962 (MS medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA and 2.69 ?M ?-naphthalene acetic acid (NAA, or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.

  20. TRANSFORMER

    Science.gov (United States)

    Baker, W.R.

    1959-08-25

    Transformers of a type adapted for use with extreme high power vacuum tubes where current requirements may be of the order of 2,000 to 200,000 amperes are described. The transformer casing has the form of a re-entrant section being extended through an opening in one end of the cylinder to form a coaxial terminal arrangement. A toroidal multi-turn primary winding is disposed within the casing in coaxial relationship therein. In a second embodiment, means are provided for forming the casing as a multi-turn secondary. The transformer is characterized by minimized resistance heating, minimized external magnetic flux, and an economical construction.

  1. Cross-correlation of bio-signals using continuous wavelet transform and genetic algorithm.

    Science.gov (United States)

    Sukiennik, Piotr; Białasiewicz, Jan T

    2015-05-30

    Continuous wavelet transform allows to obtain time-frequency representation of a signal and analyze short-lived temporal interaction of concurrent processes. That offers good localization in both time and frequency domain. Scalogram and coscalogram analysis of two signal interaction dynamics gives an indication of the cross-correlation of analyzed signals in both domains. We have used genetic algorithm with a fitness function based on signals convolution to find time delay between investigated signals. Two methods of cross-correlation are proposed: one that finds single delay for analyzed signals, and one returns a vector of delay values for each of wavelet transform sub-band center frequencies. Algorithms were implemented using MATLAB. We have extracted the data of simultaneously recorded encephalogram and arterial blood pressure and have investigated their interaction dynamics. We found time delay whose value cannot be precisely determined by scalograms and coscalogram inspection. The biomedical signals used come from MIMIC database. Cross-correlation of two complex signals is commonly performed using fast Fourier transform. It works well for signals with invariant frequency content. We have determined the time delay between analyzed signals using wavelet scalograms and we have accordingly shifted one of them, aligning associated events. Their coscalogram indicates the cross-correlation of the associated events. Introducing new methods of wavelet transform in cross-correlation analysis has proven to be beneficial to the gain of the information about process interaction. Introduced solutions could be used to reason about causality between processes and gain bigger insight regarding analyzed systems. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. A comparison of the phenotypic and genetic stability of recombinant Trichoderma spp. generated by protoplast- and Agrobacterium-mediated transformation.

    Science.gov (United States)

    Cardoza, Rosa Elena; Vizcaino, Juan Antonio; Hermosa, Maria Rosa; Monte, Enrique; Gutiérrez, Santiago

    2006-08-01

    Four different Trichoderma strains, T. harzianum CECT 2413, T. asperellum T53, T. atroviride T11 and T. longibrachiatum T52, which represent three of the four sections contained in this genus, were transformed by two different techniques: a protocol based on the isolation of protoplasts and a protocol based on Agrobacterium-mediated transformation. Both methods were set up using hygromycin B or phleomycin resistance as the selection markers. Using these techniques, we obtained phenotypically stable transformants of these four different strains. The highest transformation efficiencies were obtained with the T. longibrachiatum T52 strain: 65-70 transformants/microg DNA when transformed with the plasmid pAN7-1 (hygromycin B resistance) and 280 transformants/107 spores when the Agrobacterium-mediated transformation was performed with the plasmid pUR5750 (hygromycin B resistance). Overall, the genetic analysis of the transformants showed that some of the strains integrated and maintained the transforming DNA in their genome throughout the entire transformation and selection process. In other cases, the integrated DNA was lost.

  3. [Construction of an integration vector carrying hygromycin B resistance gene and its genetic transformation in Rhizopus oryzae].

    Science.gov (United States)

    Zhang, Min; Jiang, Shaotong; Zheng, Juan; Zheng, Zhi; Li, Xingjiang; Pan, Lijun; Luo, Shuizhong

    2015-08-01

    To construct a system of genetic transformation suitable for Rhizopus oryzae, we constructed a single-exchange vector pBS-hygro carrying hygromycin B resistance gene (hph) as its selective marker using gene splicing by overlap extension PCR (SOE PCR) technique. We introduced this recombinant vector into Rhizopus oryzae AS 3.819 by PEG/CaCl2-mediated transformation of protoplast, electroporation of protoplast and germinated spores; and we studied the effects of hydrolysis time, field strength and spore germination time on transformation frequency. We conducted quantitative real-time PCR (qPCR) assay to determine the gene copy number of ldhA integrated in the genome of R. oryzae transformants and its effect on the stability of transformants. We successfully achieved R. oryzae transformants integrated with pBS-hygro-ldhA vector. The optimal hydrolysis time for protoplast production was 140 min, and the optimal field strength of electroporation pulse for protoplast was 13 kV/cm. The optimal germination time of spores for electroporation was 2.5 h, and the optimal field strength of electroporation pulse was 14 kV/cm. The transformation frequency of method based on germinated spores was generally higher than the methods based on protoplast. The qPCR test results suggested that transformants with high copy number of integration in a certain range were relatively stable. Our results provided basis and support for metabolic regulation and genetic engineering breeding of R. oryzae.

  4. Performance Optimization of Discrete Wavelets Transform Based Image Watermarking Using Genetic Algorithms

    Directory of Open Access Journals (Sweden)

    A. Al-Haj

    2008-01-01

    Full Text Available The excellent spatial localization, frequency spread and multi-resolution characteristics of the Discrete Wavelets Transform (DWT, which were similar to the theoretical models of the human visual system, facilitated the development of many imperceptible and robust DWT-based watermarking algorithms. There had been extremely few proposed algorithms on optimized DWT-based image watermarking that can simultaneously provide perceptual transparency and robustness since these two watermarking requirements are conflicting, in this study we treat the DWT-based image watermarking problem as an optimization problem and solve it using genetic algorithms. We demonstrate through the experimental results we obtained that optimal DWT-based image watermarking can be achieved only if watermarking has been applied at specific wavelet sub-bands and by using specific watermark-amplification values.

  5. Stable genetic transformation of Jatropha curcas via Agrobacterium tumefaciens-mediated gene transfer using leaf explants

    KAUST Repository

    Kumar, Nitish

    2010-07-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel crop. A simple and reproducible protocol was developed for Agrobacterium tumefaciens-mediated stable genetic transformation of J. curcas using leaf explains. Agrobacterium strain LBA 4404 harbouring the binary vector pCAMBIA 1304 having sense-dehydration responsive element binding (S-DREB2A), beta-glucuronidase (gus), and hygromycin-phosphotransferase (hpt) genes were used for gene transfer. A number of parameters such as preculture of explains, wounding of leaf explants, Agrobacterium growth phase (OD), infection duration, co-cultivation period, co-cultivation medium pH, and acetosyringone, were studied to optimized transformation efficiency. The highest transformation efficiency was achieved using 4-day precultured, non-wounded leaf explants infected with Agrobacterium culture corresponding to OD(600)=0.6 for 20 min, followed by co-cultivation for 4 days in a co-cultivation medium containing 100 mu M acetosyringone, pH 5.7. Co-cultivated leaf explants were initially cultured on Murashige and Skoog (MS) medium supplemented with 2.27 mu M thidiazuron (TDZ) for regeneration of shoot buds, followed by selection on same medium with 5 mu g ml(-1) hygromycin. Selected shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for proliferation. The proliferated shoots were elongated on MS medium supplemented with 2.25 mu M BA and 8.5 mu M indole-3-acetic acid (IAA). The elongated shoots were rooted on half strength MS medium supplemented with 15 mu M indole-3-butyric acid (IBA), 5.7 mu M IAA, 5.5 mu M NAA, and 0.25 mg l(-1) activated charcoal. GUS histochemical analysis of the transgenic tissues further confirmed the transformation event. PCR and DNA gel blot hybridization were performed to confirm the presence of transgene. A transformation efficiency of 29% was

  6. Genetic structure of avian myeloblastosis virus, released from transformed myeloblasts as a defective virus particle

    Science.gov (United States)

    Duesberg, Peter H.; Bister, Klaus; Moscovici, Carlo

    1980-01-01

    Chicken myeloblasts transformed by avian myeloblastosis virus (AMV) in the absence of nondefective helper virus (termed nonproducer cells) were found to release a defective virus particle (DVP) that contains avian tumor viral gag proteins but lacks envelope glycoprotein and a DNA polymerase. Nonproducer cells contain a Pr76 gag precursor protein and also a protein that is indistinguishable from the Pr180 gag-pol protein of nondefective viruses. The RNA of the DVP is 7.5 kilobases (kb) long and is 0.7 kb shorter than the 8.2-kb RNAs of the helper viruses of AMV, MAV-1 and MAV-2. Comparisons based on RNA·cDNA hybridization and mapping of RNase T1-resistant oligonucleotides indicated that DVP RNA shares with MAV RNAs nearly isogenic 5′-terminal gag and pol-related sequences of 5.3 kb and a 3′-terminal c-region of 0.7 kb that is different from that found in other avian tumor viruses. Adjacent to the c-region, DVP RNA contains a contiguous specific sequence of 1.5 kb defined by 14 specific oligonucleotides. Except for two of these oligonucleotides that map at its 5′ end, this sequence is unrelated to any sequences of nondefective avian tumor viruses of four different envelope subgroups as well as to the specific sequences of fibroblast-transforming avian acute leukemia and sarcoma viruses of four different RNA subgroups. The specific sequence of the DVP RNA is present in infectious stocks of AMV from this and other laboratories in an AMV-transformed myeloblast line from another laboratory, and it is about 70% related to nucleotide sequences of E26 virus, an independent isolate of an AMV-like virus. Preliminary experiments show DVP to be leukemogenic if fused into susceptible cells in the presence of helper virus. We conclude that DVP RNA is the leukemogenic component of infectious AMV and that its specific sequence, termed AMV, may carry genetic information for oncogenicity. Thus we have found here a transformation-specific RNA sequence, unrelated to helper virus

  7. Genetic transformation via Agrobacterium tumefaciens in embryogenic cell suspensions of the plantain hybrid cultivar FHIA 21 (AAAB

    Directory of Open Access Journals (Sweden)

    Boris Chong

    2002-01-01

    Full Text Available The genetic improvement of plantain and banana is of great importance for its level of consumption on a world-wide scale. Genetic transformation constitutes one alternative for genetic improvement and has complemented traditional techniques. In this work some parameters of genetic transformation of plantain were studied by means of transient expression of â-glucoronidase in the hybrid cultivar FHIA-21(AAAB by Agrobacterium tumefaciens. A study with the β-Agrobacterium tumefaciens strains AT-2260 and EHA-105, both with the plasmid pCAMBIA-3301 was done. A comparison between the time of infection and time of co-culture was studied. The strain of better behavior was EHA-105. In the study of the time of infection and co-culture, the best combination resulted to be the one with two hours of infection and six days of co-culture. Key words: At-2260, EHA-105, β-glucoronidase, Musa

  8. Transport and transformation of genetic information in the critical zone: The case of antibiotic resistance genes

    Science.gov (United States)

    Zhu, Y. G.

    2015-12-01

    In addition to material and energy flows, the dynamics and functions of the Earth's critical zone are intensively mediated by biological actions performed by diverse organisms. These biological actions are modulated by the expression of functional genes and their translation into enzymes that catalyze geochemical reactions, such as nutrient turnover and pollutant biodegradation. Although geobiology, as an interdisciplinary research area, is playing and vital role in linking biological and geochemical processes at different temporal and spatial scales, the distribution and transport of functional genes have rarely been investigated from the Earth's critical zone perspectives. To illustrate the framework of studies on the transport and transformation of genetic information in the critical zone, antibiotic resistance is taken as an example. Antibiotic resistance genes are considered as a group of emerging contaminants, and their emergence and spread within the critical zone on one hand are induced by anthropogenic activities, and on other hand are threatening human health worldwide. The transport and transformation of antibiotic resistance genes are controlled by both horizontal gene transfer between bacterial cells and the movement of bacteria harboring antibiotic resistance genes. In this paper, the fate and behavior of antibiotic resistance genes will be discussed in the following aspects: 1) general overview of environmental antibiotic resistance; 2) high through quantification of the resistome in various environmental media; 3) pathways of resistance gene flow within the critical zone; and 4) potential strategies in mitigating antibiotic resistance, particularly from the critical zone perspectives.

  9. Genetic transformation via somatic embryogenesis to establish herbicide-resistant opium poppy.

    Science.gov (United States)

    Facchini, Peter J; Loukanina, Natalia; Blanche, Vincent

    2008-04-01

    A reliable genetic transformation protocol via somatic embryogenesis has been developed for the production of fertile, herbicide-resistant opium poppy plants. Transformation was mediated by Agrobacterium tumefaciens using the pCAMBIA3301 vector, which harbors the phosphinothricin acetyltransferase (pat) gene driven by a tandem repeat of the cauliflower mosaic virus (CaMV) 35S promoter and the beta-glucuronidase (gus) structural gene driven by a single copy of the CaMV 35S promoter between left- and right-border sequences. Co-cultivation of explants and A. tumefaciens was performed in the presence of 50 microM ATP and 50 microM MgCl(2). Root explants pre-cultured on callus induction medium were used for transformation. Herbicide-resistant, proliferating callus was obtained from explants on a medium containing both 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA). Globular embryogenic callus, induced by removal of the BA from the medium, was placed on a hormone-free medium to form somatic embryos, which were converted to plantlets under specific culture conditions. Plantlets with roots were transferred to soil, allowed to mature and set seed. Both pat and gus gene transcripts, and PAT and GUS enzyme activities were detected in the transgenic lines tested. Histochemical localization of GUS activity in T(1) opium poppy plants revealed transgene expression in most tissues of all plant organs. The protocol required 8-12 months to establish transgenic T(1) seed stocks and was developed using a commercial opium poppy cultivar that produces high levels of pharmaceutical alkaloids.

  10. Agrobacterium-mediated genetic transformation of Coffea arabica (L. is greatly enhanced by using established embryogenic callus cultures

    Directory of Open Access Journals (Sweden)

    Lashermes Philippe

    2011-05-01

    Full Text Available Abstract Background Following genome sequencing of crop plants, one of the main challenges today is determining the function of all the predicted genes. When gene validation approaches are used for woody species, the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. Embryogenic calli have frequently been the target tissue for transformation, but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants, including Coffea arabica. Results We identified the conditions required for successful long-term proliferation of embryogenic cultures in C. arabica and designed a highly efficient and reliable Agrobacterium tumefaciens-mediated transformation method based on these conditions. The transformation protocol with LBA1119 harboring pBin 35S GFP was established by evaluating the effect of different parameters on transformation efficiency by GFP detection. Using embryogenic callus cultures, co-cultivation with LBA1119 OD600 = 0.6 for five days at 20 °C enabled reproducible transformation. The maintenance conditions for the embryogenic callus cultures, particularly a high auxin to cytokinin ratio, the age of the culture (optimum for 7-10 months of proliferation and the use of a yellow callus phenotype, were the most important factors for achieving highly efficient transformation (> 90%. At the histological level, successful transformation was related to the number of proembryogenic masses present. All the selected plants were proved to be transformed by PCR and Southern blot hybridization. Conclusion Most progress in increasing transformation efficiency in coffee has been achieved by optimizing the production conditions of embryogenic cultures used as target tissues for transformation. This is the first time that a strong positive effect of the age of the culture on transformation efficiency was demonstrated. Our

  11. A genetic-algorithm-based method to find unitary transformations for any desired quantum computation and application to a one-bit oracle decision problem

    Science.gov (United States)

    Bang, Jeongho; Yoo, Seokwon

    2014-12-01

    We propose a genetic-algorithm-based method to find the unitary transformations for any desired quantum computation. We formulate a simple genetic algorithm by introducing the "genetic parameter vector" of the unitary transformations to be found. In the genetic algorithm process, all components of the genetic parameter vectors are supposed to evolve to the solution parameters of the unitary transformations. We apply our method to find the optimal unitary transformations and to generalize the corresponding quantum algorithms for a realistic problem, the one-bit oracle decision problem, or the often-called Deutsch problem. By numerical simulations, we can faithfully find the appropriate unitary transformations to solve the problem by using our method. We analyze the quantum algorithms identified by the found unitary transformations and generalize the variant models of the original Deutsch's algorithm.

  12. A genetic-algorithm-based method to find unitary transformations for any desired quantum computation and application to a one-bit oracle decision problem

    Energy Technology Data Exchange (ETDEWEB)

    Bang, Jeongho [Seoul National University, Seoul (Korea, Republic of); Hanyang University, Seoul (Korea, Republic of); Yoo, Seokwon [Hanyang University, Seoul (Korea, Republic of)

    2014-12-15

    We propose a genetic-algorithm-based method to find the unitary transformations for any desired quantum computation. We formulate a simple genetic algorithm by introducing the 'genetic parameter vector' of the unitary transformations to be found. In the genetic algorithm process, all components of the genetic parameter vectors are supposed to evolve to the solution parameters of the unitary transformations. We apply our method to find the optimal unitary transformations and to generalize the corresponding quantum algorithms for a realistic problem, the one-bit oracle decision problem, or the often-called Deutsch problem. By numerical simulations, we can faithfully find the appropriate unitary transformations to solve the problem by using our method. We analyze the quantum algorithms identified by the found unitary transformations and generalize the variant models of the original Deutsch's algorithm.

  13. Genetic Transformation of Wheat (Triticum aestivum L):A Review%小麦遗传转化研究进展

    Institute of Scientific and Technical Information of China (English)

    Abdul Razzaq; 马峙英; 王海波

    2004-01-01

    Gradual progress made in genetic transformation of wheat is presented in this paper. Information on promoters, antibiotic, herbicide and auxotrophic markers, and various traits of wheat modified through genetic transformation, is provided. In addition the methods used for wheat transformation are discussed. Though significant efforts have been made for genetic transformation of wheat mainly through particle bombardment method but transformation efficiency is still low for mass production of fertile transgenic plants. Studies on the inheritance of transgenes and its incorporation into commercial elite cultivars are not significant. Agrobacterium mediated transformation seems to have better prospects for wheat transformation in future due to its advantages over particle bombardment. In planta transformation of wheat tissues seems possible only with A grobacterium.

  14. Agrobacterium-mediated genetic transformation of yam (Dioscorea rotundata: an important tool for functional study of genes and crop improvement

    Directory of Open Access Journals (Sweden)

    Evans eNyaboga

    2014-09-01

    Full Text Available Although genetic transformation of clonally propagated crops has been widely studied as a tool for crop improvement and as a vital part of the development of functional genomics resources, there has been no report of any existing Agrobacterium-mediated transformation of yam (Dioscorea spp. with evidence of stable integration of T-DNA. Yam is an important crop in the tropics and subtropics providing food security and income to over 300 million people. However, yam production remains constrained by increasing levels of field and storage pests and diseases. A major constraint to the development of biotechnological approaches for yam improvement has been the lack of an efficient and robust transformation and regeneration system. In this study, we developed an Agrobacterium-mediated transformation of Dioscorea rotundata using axillary buds as explants. Two cultivars of D. rotundata were transformed using Agrobacterium tumefaciens harboring the binary vectors containing selectable marker and reporter genes. After selection with appropriate concentrations of antibiotic, shoots were developed on shoot induction and elongation medium. The elongated antibiotic-resistant shoots were subsequently rooted on medium supplemented with selection agent. Successful transformation was confirmed by PCR, Southern blot analysis and reporter genes assay. Expression of gusA gene in transgenic plants was also verified by RT-PCR analysis. Transformation efficiency varied from 9.4% to 18.2% depending on the cultivars, selectable marker genes and the Agrobacterium strain used for transformation. It took 3–4 months from Agro-infection to regeneration of complete transgenic plant. Here we report an efficient, fast and reproducible protocol for Agrobacterium-mediated transformation of D. rotundata using axillary buds as explants, which provides a useful platform for future genetic engineering studies in this economically important crop.

  15. Agrobacterium-mediated genetic transformation of yam (Dioscorea rotundata): an important tool for functional study of genes and crop improvement.

    Science.gov (United States)

    Nyaboga, Evans; Tripathi, Jaindra N; Manoharan, Rajesh; Tripathi, Leena

    2014-01-01

    Although genetic transformation of clonally propagated crops has been widely studied as a tool for crop improvement and as a vital part of the development of functional genomics resources, there has been no report of any existing Agrobacterium-mediated transformation of yam (Dioscorea spp.) with evidence of stable integration of T-DNA. Yam is an important crop in the tropics and subtropics providing food security and income to over 300 million people. However, yam production remains constrained by increasing levels of field and storage pests and diseases. A major constraint to the development of biotechnological approaches for yam improvement has been the lack of an efficient and robust transformation and regeneration system. In this study, we developed an Agrobacterium-mediated transformation of Dioscorea rotundata using axillary buds as explants. Two cultivars of D. rotundata were transformed using Agrobacterium tumefaciens harboring the binary vectors containing selectable marker and reporter genes. After selection with appropriate concentrations of antibiotic, shoots were developed on shoot induction and elongation medium. The elongated antibiotic-resistant shoots were subsequently rooted on medium supplemented with selection agent. Successful transformation was confirmed by polymerase chain reaction, Southern blot analysis, and reporter genes assay. Expression of gusA gene in transgenic plants was also verified by reverse transcription polymerase chain reaction analysis. Transformation efficiency varied from 9.4 to 18.2% depending on the cultivars, selectable marker genes, and the Agrobacterium strain used for transformation. It took 3-4 months from Agro-infection to regeneration of complete transgenic plant. Here we report an efficient, fast and reproducible protocol for Agrobacterium-mediated transformation of D. rotundata using axillary buds as explants, which provides a useful platform for future genetic engineering studies in this economically important

  16. Conjugative plasmid transfer from Escherichia coli is a versatile approach for genetic transformation of thermophilic Bacillus and Geobacillus species.

    Science.gov (United States)

    Tominaga, Yurie; Ohshiro, Takashi; Suzuki, Hirokazu

    2016-05-01

    We previously demonstrated efficient transformation of the thermophile Geobacillus kaustophilus HTA426 using conjugative plasmid transfer from Escherichia coli BR408. To evaluate the versatility of this approach to thermophile transformation, this study examined genetic transformation of various thermophilic Bacillus and Geobacillus spp. using conjugative plasmid transfer from E. coli strains. E. coli BR408 successfully transferred the E. coli-Geobacillus shuttle plasmid pUCG18T to 16 of 18 thermophiles with transformation efficiencies between 4.1 × 10(-7) and 3.8 × 10(-2)/recipient. Other E. coli strains that are different from E. coli BR408 in intracellular DNA methylation also generated transformants from 9 to 15 of the 18 thermophiles, including one that E. coli BR408 could not transform, although the transformation efficiencies of these strains were generally lower than those of E. coli BR408. The conjugation was performed by simple incubation of an E. coli donor and a thermophile recipient without optimization of experimental conditions. Moreover, thermophile transformants were distinguished from abundant E. coli donor only by high temperature incubation. These observations suggest that conjugative plasmid transfer, particularly using E. coli BR408, is a facile and versatile approach for plasmid introduction into thermophilic Bacillus and Geobacillus spp., and potentially a variety of other thermophiles.

  17. Control of competence for DNA transformation in streptococcus suis by genetically transferable pherotypes.

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    Edoardo Zaccaria

    Full Text Available Here we show that S. suis, a major bacterial pathogen of pigs and emerging pathogen in humans responds to a peptide pheromone by developing competence for DNA transformation. This species does not fall within any of the phylogenetic clusters of streptococci previously shown to regulate competence via peptide pheromones suggesting that more species of streptococci may be naturally competent. Induction of competence was dependent on ComX, a sigma factor that controls the streptococcal late competence regulon, extracellular addition of a comX-inducing peptide (XIP, and ComR, a regulator of comX. XIP was identified as an N-terminally truncated variant of ComS. Different comS alleles are present among strains of S. suis. These comS alleles are not functionally equivalent and appear to operate in conjuction with a cognate ComR to regulate comX through a conserved comR-box promoter. We demonstrate that these 'pherotypes' can be genetically transferred between strains, suggesting that similar approaches might be used to control competence induction in other lactic acid bacteria that lack ComR/ComS homologues but possess comX and the late competence regulon. The approaches described in this paper to identify and optimize peptide-induced competence may also assist other researchers wishing to identify natural competence in other bacteria. Harnessing natural competence is expected to accelerate genetic research on this and other important streptococcal pathogens and to allow high-throughput mutation approaches to be implemented, opening up new avenues for research.

  18. Organogenesis and plant selection by kanamycin of strawberry in vitro aiming at genetic transformation

    Directory of Open Access Journals (Sweden)

    Letícia Reis

    2015-08-01

    Full Text Available The success of genetic transformation of the strawberry (Fragaria x ananassa is strongly influenced by the cultivar and depends on efficient regeneration and plant-transformed selection. In plant regeneration, efficient organogenesis depends on the concentrations of plant growth regulators that are present in the culture medium, and the response varies according to the explant type and cultivar. Similarly, in the selection of transformed plants, the sensitivity to selective agents induced by marker genes depends on the concentration of the selective substance in the regeneration medium and must be adjusted according to the chosen cultivar. The aim of this work was to test different combinations and concentrations of TDZ (Thidiazuron and IBA (indolebutyric-acid added to the base medium MS (MURASHIGE; SKOOG, 1962 for shoot regeneration of Camiño Real and Festival cultivars from leaf and petiole explants. In the case of ‘Camiño Real’, 1.0, 1.5 and 2.0 mg L-1 of TDZ combined with 0.1, 0.3 and 0.5 mg L-1 of IBA were tested. In the case of ‘Festival’, 1.0, 1.5 and 2.0 mg L-1 of TDZ combined with 0.1 and 0.3 mg L-1 of IBA were evaluated. Another aim was to define the lowest concentration of the selective agent Kanamycin (marker gene npt II able to inhibit regeneration of the untransformed shoots. For this purpose, Kanamycin doses of 0, 2, 4, 6, 8 and 10 mg L-1 were added to the pre-established regeneration medium. In the petiole explant test, normal shoot regeneration did not occur; thus, only shoot regeneration and the sensitivity to Kanamycin from leaf explants were analyzed. For ‘Camiño Real’ the best shoot regeneration was established with a combination of 2.0 mg L-1 of TDZ + 0.1 mg L-1 of IBA. On the other hand, for ‘Festival’ the best shoot regeneration was established with a combination of 2.5 mg L-1 of TDZ + 0.1 mg L-1 of IBA. As far as Kanamycin sensitivity is concerned, total inhibition of the shoots was observed with 10 mg

  19. Genetic transformation of Drosophila willistoni using piggyBac transposon and GFP

    Directory of Open Access Journals (Sweden)

    Manuela Finokiet

    2007-01-01

    Full Text Available Studies were carried out on the use of piggyBac transposable element as vector and the green fluorescent protein (EGFP from the jellyfish, Aquorea victoria, as a genetic marker for the transformation of Drosophila willistoni. Preblastoderm embryos of D. willistoni white mutant were microinjected with a plasmid containing the EGFP marker and the piggyBac ITRs, together with a helper plasmid containing the piggyBac transposase placed under the control of the D. melanogaster hsp70 promoter. G0 adults transformants were recovered at a frequency of approximately 67%. Expression of EGFP in larvae, pupae and adults was observed up to the third generation, suggesting that this transposon was not stable in D. willistoni. Transformed individuals displayed high levels of EGFP expression during larvae and adult stages in the eye, abdomen, thorax and legs, suggesting a wide expression pattern in this species than reported to other species of Drosophilidae.Descrevemos neste trabalho a transformação genética de Drosophila willistoni empregando o elemento transponível piggyBac como vetor e o gene EGFP (green fluorescent protein retirado da água-viva Aquorea victoria, como marcador de transformação. Embriões de D. willistoni em estágio pré-blastoderme, mutantes para o gene white, foram microinjetados com plasmídio contendo o marcador EGFP e as regiões ITRs do transposon piggyBac concomitantemente com um plasmídio auxiliar possuindo o gene da transposase de piggyBac sobre o controle do promotor do gene hsp70 de Drosophila melanogaster. Adultos transformantes Go foram gerados em uma taxa de 67%. A expressão de GFP em larvas, pupas e adultos foi observada somente até a terceira geração, sugerindo que este transposon não é estável em D. willistoni. Os indivíduos transformados exigem um alto nível de expressão de EGFP durante os estágios de larva e, também em adultos o gene marcador é expresso nos olhos, abdome, tórax e patas, mostrando um

  20. Genetic Transformation and Genomic Resources for Next-Generation Precise Genome Engineering in Vegetable Crops

    Science.gov (United States)

    Cardi, Teodoro; D’Agostino, Nunzio; Tripodi, Pasquale

    2017-01-01

    In the frame of modern agriculture facing the predicted increase of population and general environmental changes, the securement of high quality food remains a major challenge to deal with. Vegetable crops include a large number of species, characterized by multiple geographical origins, large genetic variability and diverse reproductive features. Due to their nutritional value, they have an important place in human diet. In recent years, many crop genomes have been sequenced permitting the identification of genes and superior alleles associated with desirable traits. Furthermore, innovative biotechnological approaches allow to take a step forward towards the development of new improved cultivars harboring precise genome modifications. Sequence-based knowledge coupled with advanced biotechnologies is supporting the widespread application of new plant breeding techniques to enhance the success in modification and transfer of useful alleles into target varieties. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 system, zinc-finger nucleases, and transcription activator-like effector nucleases represent the main methods available for plant genome engineering through targeted modifications. Such technologies, however, require efficient transformation protocols as well as extensive genomic resources and accurate knowledge before they can be efficiently exploited in practical breeding programs. In this review, we revise the state of the art in relation to availability of such scientific and technological resources in various groups of vegetables, describe genome editing results obtained so far and discuss the implications for future applications. PMID:28275380

  1. Genetic transformation of mature embryos of bread (T. aestivum) and pasta (T. durum) wheat genotypes.

    Science.gov (United States)

    Moghaieb, Reda E A; El-Arabi, Nagwa I; Momtaz, Osama A; Youssef, Sawsan S; Soliman, Mohamed H

    2010-01-01

    The objective of the present study is to develop an efficient protocol for regeneration of transgenic wheat plants using Agrobacterium- mediated transformation of mature embryos of hexaploid bread wheat (Triticum aestivum) and tetraploid pasta wheat (Triticum durum). The data indicated that embryogenic calli were formed within 7 days in the presence of 2 mgl-1 2,4-D. Adventitious shoots emerged from the embryonic calli in the presence of 2 mgl-1 BA. Shoot regeneration frequency varied between wheat cultivars according to their genetic background differences. Regeneration frequency was higher in the cultivar Gemmiza 10 (95 %) compared with the other cultivars tested. Mature embryos derived callus of the cultivars Gemmiza 10 and Gemmiza 9 were co-cultivated with A. tumefaciens strain LBA4404 harboring a binary vector pBI-121 containing the neomycin phosphotransferase-II gene (npt-II). The resulted putative transgenic plantlets were able to grow on kanamycin containing medium. A successful integration of the transgene was confirmed by analyzing the T0 plantlets using Southern hybridization and PCR amplification. The gus gene expression can be detected only in the transgenic plants. The reported protocol is reproducible and can be used to regenerate transgenic wheat plants expressing the genes present in A. tumifaciens binary vectors.

  2. Histological study of organogenesis in Cucumis melo L. after genetic transformation: why is it difficult to obtain transgenic plants?

    Science.gov (United States)

    Chovelon, V; Restier, V; Giovinazzo, N; Dogimont, C; Aarrouf, J

    2011-11-01

    Melon (Cucumis melo L.) is widely considered as a recalcitrant species for genetic transformation. In this study, we developed different regeneration and transformation protocols and we examined the regeneration process at different steps by histological studies. The highest regeneration rate (1.13 ± 0.02 plants per explant) was obtained using cotyledon explants of the 'Védrantais' genotype on Murashige and Skoog (MS) medium supplemented with 0.2 mg/l 6-benzylaminopurine (BAP) and 0.2 mg/l dimethylallylaminopurine (2-iP). Agrobacterium tumefaciens-mediated transformations with the uidA reporter gene were realized on cotyledon explants cultivated in these conditions: 70-90% of explants expressed a transient GUS activity during the early stages of regeneration, however, only few transgenic plants were obtained (1.8-4.5% of stable transformation with the GV2260pBI101 strain). These results revealed a low capacity of melon GUS-positive cells to regenerate transgenic plants. To evaluate the influence of the Agrobacterium infection on plant regeneration, histological analyses were conducted on explants 2, 7, 15, and 28 days after co-culture with the GV2260pBI101 strain. Genetic transformation occurred in epidermal and sub-epidermal cells and reached the meristematic structures expressing a high level of GUS activity during 14 days of culture; but after this period, most of the meristematic structures showed premature cell vacuolization and disorganization. This disruption of the GUS-positive meristematic areas could be responsible of the difficulties encountered to regenerate melon plants after genetic transformation.

  3. Genetic transformation of the plant pathogens Phytophthora capsici and Phytophthora parasitica.

    OpenAIRE

    Bailey, A. M.; Mena, G L; Herrera-Estrella, L

    1991-01-01

    Phytophthora capsici and P.parasitica were transformed to hygromycin B resistance using plasmids pCM54 and pHL1, which contain the bacterial hygromycin B phosphotransferase gene (hph) fused to promoter elements of the Ustilago maydis heat shock hsp70 gene. Enzymes Driselase and Novozyme 234 were used to generate protoplasts which were then transformed following exposure to plasmid DNA and polyethylene glycol 6000. Transformation frequencies of over 500 transformants per micrograms of DNA per ...

  4. Novel recombinant binary vectors harbouring Basta (bar) gene as a plant selectable marker for genetic transformation of plants.

    Science.gov (United States)

    Nada, Reham M

    2016-04-01

    Genetic transformation is one of the most widely used technique in crop improvement. However, most of the binary vectors used in this technique, especially cloning based, contain antibiotic genes as selection marker that raise serious consumer and environmental concerns; moreover, they could be transferred to non-target hosts with deleterious effects. Therefore, the goal of this study was reconstruction of the widely used pBI121 binary vector by substituting the harmful antibiotic selection marker gene with a less-harmful selection marker, Basta (herbicide resistance gene). The generated vectors were designated as pBI121NB and pBI121CB, in which Basta gene was expressed under the control of Nos or CaMV 35S promoter, respectively. The successful integration of the new inserts into both the vectors was confirmed by PCR, restriction digestion and sequencing. Both these vectors were used in transforming Arabidopsis, Egyptian wheat and barley varieties using LBA4404 and GV3101 Agrobacterium strains. The surfactant Tween-20 resulted in an efficient transformation and the number of Arabidopsis transformants was about 6-9 %. Soaked seeds of wheat and barley were transformed with Agrobacterium to introduce the bacteria to the growing shoot apices. The percentage of transgenic lines was around 16-17 and 14-15 % for wheat and barley, respectively. The quantitative studies presented in this work showed that both LBA4404 and GV3101 strains were suitable for transforming Egyptian wheat and barley.

  5. Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter

    Directory of Open Access Journals (Sweden)

    Lísia Borges Attílio

    2013-07-01

    Full Text Available The objective of this work was to produce transgenic 'Pêra' and 'Valência' sweet orange plants using the D4E1 gene driven by the Arabidopsis thaliana phloem protein (AtPP2 promoter and to quantify transgene expression in different transformation events. Genetic transformation experiments were carried out with epicotyl segments co‑cultivated with Agrobacterium tumefaciens. Six plants from 'Pêra' sweet orange and seven plants from 'Valência' sweet orange were confirmed as different transgenic events by means of the polymerase chain reaction (PCR and the Southern blot techniques. Transgene expression was quantified using real‑time quantitative PCR. D4E1 gene expression levels vary from 5 up to 50 times among different transformation events.

  6. An efficient method for Agrobacterium-mediated genetic transformation and plant regeneration in cumin (Cuminum cyminum L.).

    Science.gov (United States)

    Pandey, Sonika; Mishra, Avinash; Patel, Manish Kumar; Jha, Bhavanath

    2013-09-01

    Cumin is an annual herbaceous medicinally important plant having diverse applications. An efficient and reproducible method of Agrobacterium-mediated genetic transformation was herein established for the first time. A direct regeneration method without callus induction was optimised using embryos as explant material in Gamborg's B5 medium supplemented with 0.5-μM 6-benzyladenine and 2.0-μM α-naphthalene acetic acid. About 1,020 embryos (a mean of 255 embryos per batch) were used for the optimisation of transformation conditions. These conditions were an Agrobacterium cell suspension of 0.6 OD600, a co-cultivation time of 72 h, 300-μM acetosyringone and wounding of explants using a razor blade. Pre-cultured elongated embryos were treated using optimised conditions. About 720 embryos (a mean of 180 embryos per batch) were used for transformation and 95 % embryos showed transient β-glucuronidase expression after co-cultivation. Putative transformed embryos were cultured on B5 medium for shoot proliferation and 21 regenerated plants were obtained after selection and allowed to root. T0 plantlets showed β-glucuronidase expression and gene integration was confirmed via PCR amplification of 0.96 and 1.28 kb fragments of the hygromycin-phosphotransferase II and β-glucuronidase genes, respectively. In this study, a transformation efficiency of 1.5 % was demonstrated and a total of 11 transgenic plants were obtained at the hardening stage, however, only four plants acclimatised during hardening. Gene copy number was analysed by Southern blot analysis of hardened plants and single-copy gene integration was observed. This is the first successful attempt of Agrobacterium-mediated genetic transformation of cumin.

  7. Genetic Transformation of Brassica campestris L. ssp. pekinensis via Agrobacterium- with Bt Gene

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    By means of Agrobacterium-mediated transformation, 43 kanamycin-resistant buds of Chinese cabbage were got. PCR, PCR-Southern blot and dot blot analysis were used to identify and characterize the putative transgenic plants. 26 plants had the predicted bands of the fragment of npt II gene. Insect bioassays of 4 transformants showed that toxic protein had been translated and the translation levels were different among these transformants.

  8. Genetic Transformation of Artemisia carvifolia Buch with rol Genes Enhances Artemisinin Accumulation.

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    Erum Dilshad

    Full Text Available The potent antimalarial drug artemisinin has a high cost, since its only viable source to date is Artemisia annua (0.01-0.8% DW. There is therefore an urgent need to design new strategies to increase its production or to find alternative sources. In the current study, Artemisia carvifolia Buch was selected with the aim of detecting artemisinin and then enhancing the production of the target compound and its derivatives. These metabolites were determined by LC-MS in the shoots of A. carvifolia wild type plants at the following concentrations: artemisinin (8μg/g, artesunate (2.24μg/g, dihydroartemisinin (13.6μg/g and artemether (12.8μg/g. Genetic transformation of A. carvifolia was carried out with Agrobacterium tumefaciens GV3101 harboring the rol B and rol C genes. Artemisinin content increased 3-7-fold in transgenics bearing the rol B gene, and 2.3-6-fold in those with the rol C gene. A similar pattern was observed for artemisinin analogues. The dynamics of artemisinin content in transgenics and wild type A.carvifolia was also correlated with the expression of genes involved in its biosynthesis. Real time qPCR analysis revealed the differential expression of genes involved in artemisinin biosynthesis, i.e. those encoding amorpha-4, 11 diene synthase (ADS, cytochrome P450 (CYP71AV1, and aldehyde dehydrogenase 1 (ALDH1, with a relatively higher transcript level found in transgenics than in the wild type plant. Also, the gene related to trichome development and sesquiterpenoid biosynthesis (TFAR1 showed an altered expression in the transgenics compared to wild type A.carvifolia, which was in accordance with the trichome density of the respective plants. The trichome index was significantly higher in the rol B and rol C gene-expressing transgenics with an increased production of artemisinin, thereby demonstrating that the rol genes are effective inducers of plant secondary metabolism.

  9. Genetic transformation of the plant pathogens Phytophthora capsici and Phytophthora parasitica.

    Science.gov (United States)

    Bailey, A M; Mena, G L; Herrera-Estrella, L

    1991-08-11

    Phytophthora capsici and P.parasitica were transformed to hygromycin B resistance using plasmids pCM54 and pHL1, which contain the bacterial hygromycin B phosphotransferase gene (hph) fused to promoter elements of the Ustilago maydis heat shock hsp70 gene. Enzymes Driselase and Novozyme 234 were used to generate protoplasts which were then transformed following exposure to plasmid DNA and polyethylene glycol 6000. Transformation frequencies of over 500 transformants per micrograms of DNA per 1 x 10(6) protoplasts were obtained. Plasmid pCM54 appears to be transmitted in Phytophthora spp. as an extra-chromosomal element through replication, as shown by Southern blot hybridization and by the loss of plasmid methylation. In addition, transformed strains retained their capacity of infecting Serrano pepper seedlings and Mc. Intosh apple fruits, the host plants for P.capsici and P.parasitica, respectively.

  10. Assessment of genetically modified soybean crops and different cultivars by Fourier transform infrared spectroscopy and chemometric analysis

    Directory of Open Access Journals (Sweden)

    Glaucia Braz Alcantara

    2010-06-01

    Full Text Available This paper describes the potentiality of Fourier transform infrared (FT-IR spectroscopy associated to chemometric analysis for assessment of conventional and genetically modified soybean crops. Recently, genetically modified organisms have been queried about their influence on the environment and their safety as food/feed. In this regard, chemical investigations are ever more required. Thus three different soybean cultivars distributed in transgenic Roundup ReadyTM soybean and theirs conventional counterparts were directly investigated by FT-IR spectroscopy and chemometric analysis. The application of PCA and KNN methods permitted the discrimination and classification of the genetically modified samples from conventional ones when they were separately analysed. The analyses showed the chemical variation according to genetic modification. Furthermore, this methodology was efficient for cultivar grouping and highlights cultivar dependence for discrimination between transgenic and non-transgenic samples. According to this study, FT-IR and chemometrics could be used as a quick, easy and low cost tool to assess the chemical composition variation in genetically modified organisms.

  11. Genetic transformation of the plant pathogens Phytophthora capsici and Phytophthora parasitica

    National Research Council Canada - National Science Library

    Bailey, A M; Mena, G L; Herrera-Estrella, L

    1991-01-01

    ...) fused to promoter elements of the Ustilago maydis heat shock hsp70 gene. Enzymes Driselase and Novozyme 234 were used to generate protoplasts which were then transformed following exposure to plasmid DNA and polyethylene glycol 6000...

  12. Reproduction of Meloidogyne javanica on Plant Roots Genetically Transformed by Agrobacterium rhizogenes

    OpenAIRE

    Verdejo, S.; Jaffee, B. A.; Mankau, R.

    1988-01-01

    Reproduction of Meloidogyne javanica was compared on several Agrobacterium rhizogenes-transformed root cultures under monoxenic conditions. M. javanica reproduced on all transformed roots tested; however, more females and eggs were obtained on potato and South Australian Early Dwarf Red tomato than on bindweed, Tropic tomato, lima bean, or carrot. Roots that grew at moderate rates into the agar and produced many secondary roots supported the highest reproduction. Numbers of females produced i...

  13. Identification of Moraxella bovis and related species from calves with IBK and goats by qualitative genetic transformation assay.

    Science.gov (United States)

    Kodjo, A; Exbrayat, P; Richard, Y

    1994-07-01

    Eight Moraxella bovis strains isolated from bovine pink eye, 16 Moraxella bovis related strains isolated from healthy goats nasal flora, one nonhemolytic Moraxella sp. isolated from bovine conjunctivitis and different collection strains of Moraxella and Branhamella genus were studied through the combined use of qualitative genetic transformation assay and the ability to grow on a moraxella bovis defined medium (medium MB). Crude DNA extracted from the strains studied were used to transform two mutant auxotroph competent strains of Moraxella bovis CIP 103741 and CIP 103743. Non-specific positive transformation was obtained with mutant assay strain CIP 103743 when treated with DNA from caprine or bovine Moraxella strains, collection strains of Moraxella bovis, Moraxella lacunata, Moraxella nonliquefaciens and strains of Branhamella genus, whereas specific transformation was observed with mutant assay CIP 103741 when treated only with DNA from all the Moraxella bovis (collection and clinical isolates) and 14 of the 16 caprine Moraxella bovis related strains. The specificity and the simplicity of the test make it suitable for use in clinical laboratories.

  14. Mitochondrial Porin Isoform AtVDAC1 Regulates the Competence of Arabidopsis thaliana to Agrobacterium-Mediated Genetic Transformation

    Science.gov (United States)

    Kwon, Tackmin

    2016-01-01

    The efficiency of Agrobacterium-mediated transformation in plants depends on the virulence of Agrobacterium strains, the plant tissue culture conditions, and the susceptibility of host plants. Understanding the molecular interactions between Agrobacterium and host plant cells is crucial when manipulating the susceptibility of recalcitrant crop plants and protecting orchard trees from crown gall disease. It was discovered that Arabidopsis voltage-dependent anion channel 1 (atvdac1) mutant has drastic effects on Agrobacterium-mediated tumorigenesis and growth developmental phenotypes, and that these effects are dependent on a Ws-0 genetic background. Genetic complementation of Arabidopsis vdac1 mutants and yeast porin1-deficient strain with members of the AtVDAC gene family revealed that AtVDAC1 is required for Agrobacterium-mediated transformation, and there is weak functional redundancy between AtVDAC1 and AtVDAC3, which is independent of porin activity. Furthermore, atvdac1 mutants were deficient in transient and stable transformation by Agrobacterium, suggesting that AtVDAC1 is involved in the early stages of Agrobacterium infection prior to transferred-DNA (T-DNA) integration. Transgenic plants overexpressing AtVDAC1 not only complemented the phenotypes of the atvdac1 mutant, but also showed high efficiency of transient T-DNA gene expression; however, the efficiency of stable transformation was not affected. Moreover, the effect of phytohormone treatment on competence to Agrobacterium was compromised in atvdac1 mutants. These data indicate that AtVDAC1 regulates the competence of Arabidopsis to Agrobacterium infection. PMID:27643450

  15. Further genetic localization of the transforming sequences of the p21 v-ras gene of Harvey murine sarcoma virus

    DEFF Research Database (Denmark)

    Willumsen, B M; Ellis, R W; Scolnick, E M

    1984-01-01

    The sequences encoding the 21-kilodalton transforming protein (p21 ras) of Harvey murine sarcoma virus have previously been localized genetically to a 1.3-kilobase segment of the viral DNA (E. H. Chang, R. W. Ellis, E. M. Scolnick, and D. R. Lowy, Science 210:1249-1251, 1980). Within this segment...... of this open reading frame. By constructing a mutant of Harvey murine sarcoma virus DNA from which the first two ATG codons of this open reading frame have been deleted, we now show by transfection of the mutant viral DNA into NIH 3T3 cells that only the third ATG codon is necessary and sufficient...

  16. Evaluation of three herbicide resistance genes for use in genetic transformations and for potential crop protection in algae production.

    Science.gov (United States)

    Brueggeman, Andrew J; Kuehler, Daniel; Weeks, Donald P

    2014-09-01

    Genes conferring resistance to the herbicides glyphosate, oxyfluorfen and norflurazon were developed and tested for use as dominant selectable markers in genetic transformation of Chlamydomonas reinhardtii and as potential tools for the protection of commercial-scale algal production facilities against contamination by organisms sensitive to these broad-spectrum herbicides. A synthetic glyphosate acetyltransferase (GAT) gene, when fitted with a strong Chlamydomonas promoter, conferred a 2.7×-fold increase in tolerance to the EPSPS inhibitor, glyphosate, in transgenic cells compared with progenitor WT cells. A mutant Chlamydomonas protoporphyrinogen oxidase (protox, PPO) gene previously shown to produce an enzyme insensitive to PPO-inhibiting herbicides, when genetically engineered, generated transgenic cells able to tolerate up to 136× higher levels of the PPO inhibitor, oxyfluorfen, than nontransformed cells. Genetic modification of the Chlamydomonas phytoene desaturase (PDS) gene-based gene sequences found in various norflurazon-resistant organisms allowed production of transgenic cells tolerant to 40× higher levels of norflurazon than nontransgenic cells. The high efficiency of all three herbicide resistance genes in producing transgenic cells demonstrated their suitability as dominant selectable markers for genetic transformation of Chlamydomonas and, potentially, other eukaryotic algae. However, the requirement for high concentrations of glyphosate and its associated negative effects on cell growth rates preclude its consideration for use in large-scale production facilities. In contrast, only low doses of norflurazon and oxyfluorfen (~1.5 μm and ~0.1 μm, respectively) are required for inhibition of cell growth, suggesting that these two herbicides may prove effective in large-scale algal production facilities in suppressing growth of organisms sensitive to these herbicides.

  17. Optimization of genetic transformation of Artemisia annua L. Using Agrobacterium for Artemisinin production

    Directory of Open Access Journals (Sweden)

    Elfahmi

    2014-01-01

    Full Text Available Background: Artemisinin, a sesquiterpene lactone endoperoxide isolated from the medicinal plant Artemisia annua L., is a choice and effective drug for malaria treatment. Due to the low yield of artemisinin in plants, there is a need to enhance the production of artemisinin from A. annua and biotechnological technique may be one of the methods that can be used for the purpose. Aim: To study the transformation efficiency of Agrobacterium tumefaciens in A. annua that could be applied to enhance the production of artemisinin by means of transgenic plants. Setting and Designs: The factors influencing Agrobacterium-mediated transformation of A. annua were explored to optimize the transformation system, which included A. tumefaciens strain and effect of organosilicone surfactants. Three strains of A. tumefaciens, that is, LBA4404, GV1301, and AGL1 harboring the binary vector pCAMBIA 1303 have been used for transformation. The evaluation was based on transient β-glucuronidase (GUS. Materials and Methods: Plant cell cultures were inniatiated from the seeds of A. annua using the germination Murashige and Skoog medium. A. tumefaciens harboring pCAMBIA were tranformed into the leaves of A.annua cultures from 2-week-old-seedling and 2-month-old-seedling for 15 min by vacuum infiltration. Transformation efficiency was determinated by measuring of blue area (GUS expression on the whole leaves explant using ImageJ 1.43 software. Two organosilicon surfactants, that is, Silwet L-77 and Silwet S-408 were used to improve the transformation efficiency. Results: The transformation frequency with AGL1 strain was higher than GV3101 and LBA4404 which were 70.91, 49.25, and 45.45%, respectively. Effect of organosilicone surfactants, that is, Silwet L-77 and Silwet S-408 were tested on A. tumefaciens AGL1 and GV3101 for their level of transient expression, and on A. rhizogenes R1000 for its hairy root induction frequency. For AGL1, Silwet S-408 produced higher level of

  18. Reproduction of Meloidogyne javanica on Plant Roots Genetically Transformed by Agrobacterium rhizogenes.

    Science.gov (United States)

    Verdejo, S; Jaffee, B A; Mankau, R

    1988-10-01

    Reproduction of Meloidogyne javanica was compared on several Agrobacterium rhizogenes-transformed root cultures under monoxenic conditions. M. javanica reproduced on all transformed roots tested; however, more females and eggs were obtained on potato and South Australian Early Dwarf Red tomato than on bindweed, Tropic tomato, lima bean, or carrot. Roots that grew at moderate rates into the agar and produced many secondary roots supported the highest reproduction. Numbers of females produced in cultures of transformed potato roots increased with increasing nematode inoculum levels, whether inoculum was dispersed eggs or juveniles. Females appeared smaller, produced fewer eggs, and were found in coalesced galls at the higher inoculum levels. The ratio between the final and initial population decreased sharply as the juvenile inoculum increased. The second-stage juvenile was preferred to dispersed eggs or egg masses for inoculation of tissue culture systems because quantity and viability of inoculum were easily assessed. Meloidogyne javanica reared on transformed root cultures were able to complete their life cycles on new transformed root cultures or greenhouse tomato plants.

  19. Physiological Consequences of Genetic Transformation: Result of Target Gene Expression or Stress Reaction?

    Directory of Open Access Journals (Sweden)

    A.G. Enikeev

    2015-06-01

    Full Text Available The transgenic and non-transgenic tobacco cell cultures were analyzed for resistance to abiotic and biotic stress. The different physiological reaction of cell culture depending on T-DNA structure (or transgen structure was observed. The cell culture transformed by disarmed Agrobacterium tumefaciense A699 with pCNL 65 nptII demonstrated the same stress-resistance as non-transgenic control cell culture. The cell culture transformed by Agrobacterium tumefaciense LBA 4400 pBiCaMV nptII + hsp101 showed a raised stress-resistance to high temperature, high KF concentration, and to the action of Clavibacter michiganensis ssp sepidonicus. Obviously, the expression of transferred arabodopsis gene hsp101 provides protection properties of transgenic cell culture under the influence of various stress factors. Moreover, that agrobacterial transformation as previous stress-factor is supposed to make a contribution to formation of transgenic cell culture cross-resistance.

  20. piggybac- and PhiC31-mediated genetic transformation of the Asian tiger mosquito, Aedes albopictus (Skuse.

    Directory of Open Access Journals (Sweden)

    Geneviève M C Labbé

    Full Text Available BACKGROUND: The Asian tiger mosquito, Aedes albopictus (Skuse, is a vector of several arboviruses including dengue and chikungunya. This highly invasive species originating from Southeast Asia has travelled the world in the last 30 years and is now established in Europe, North and South America, Africa, the Middle East and the Caribbean. In the absence of vaccine or antiviral drugs, efficient mosquito control strategies are crucial. Conventional control methods have so far failed to control Ae. albopictus adequately. METHODOLOGY/PRINCIPAL FINDINGS: Germline transformation of Aedes albopictus was achieved by micro-injection of embryos with a piggyBac-based transgene carrying a 3xP3-ECFP marker and an attP site, combined with piggyBac transposase mRNA and piggyBac helper plasmid. Five independent transgenic lines were established, corresponding to an estimated transformation efficiency of 2-3%. Three lines were re-injected with a second-phase plasmid carrying an attB site and a 3xP3-DsRed2 marker, combined with PhiC31 integrase mRNA. Successful site-specific integration was observed in all three lines with an estimated transformation efficiency of 2-6%. CONCLUSIONS/SIGNIFICANCE: Both piggybac- and site-specific PhiC31-mediated germline transformation of Aedes albopictus were successfully achieved. This is the first report of Ae. albopictus germline transformation and engineering, a key step towards studying and controlling this species using novel molecular techniques and genetic control strategies.

  1. An Agrobacterium tumefaciens Strain with Gamma-Aminobutyric Acid Transaminase Activity Shows an Enhanced Genetic Transformation Ability in Plants

    Science.gov (United States)

    Nonaka, Satoko; Someya, Tatsuhiko; Zhou, Sha; Takayama, Mariko; Nakamura, Kouji; Ezura, Hiroshi

    2017-01-01

    Agrobacterium tumefaciens has the unique ability to mediate inter-kingdom DNA transfer, and for this reason, it has been utilized for plant genetic engineering. To increase the transformation frequency in plant genetic engineering, we focused on gamma-aminobutyric acid (GABA), which is a negative factor in the Agrobacterium-plant interaction. Recent studies have shown contradictory results regarding the effects of GABA on vir gene expression, leading to the speculation that GABA inhibits T-DNA transfer. In this study, we examined the effect of GABA on T-DNA transfer using a tomato line with a low GABA content. Compared with the control, the T-DNA transfer frequency was increased in the low-GABA tomato line, indicating that GABA inhibits T-DNA transfer. Therefore, we bred a new A. tumefaciens strain with GABA transaminase activity and the ability to degrade GABA. The A. tumefaciens strain exhibited increased T-DNA transfer in two tomato cultivars and Erianthus arundinacues and an increased frequency of stable transformation in tomato. PMID:28220841

  2. Stable genetic transformation of Vigna mungo L. Hepper via Agrobacterium tumefaciens.

    Science.gov (United States)

    Saini, R; Sonia; Jaiwal, P K; Jaiwal, S

    2003-06-01

    Vigna mungo is one of the large-seeded grain legumes that has not yet been transformed. We report here for the first time the production of morphologically normal and fertile transgenic plants from cotyledonary-node explants inoculated with Agrobacterium tumefaciens carrying binary vector pCAMBIA2301, the latter of which contains a neomycin phosphotransferase ( nptII) gene and a beta-glucuronidase (GUS) gene ( uidA) interrupted with an intron. The transformed green shoots, selected and rooted on medium containing kanamycin, tested positive for nptII and uidA genes by polymerase chain reaction (PCR) analysis. These shoots were established in soil and grown to maturity to collect the seeds. Mechanical wounding of the explants prior to inoculation with Agrobacterium, time lag in regeneration due to removal of the cotyledons from explants and a second round of selection at the rooting stage were found to be critical for transformation. Analysis of T(0) plants showed the expression and integration of uidA into the plant genome. GUS activity in leaves, roots, flowers, anthers and pollen grains was detected by histochemical assay. PCR analysis of T(1) progeny revealed a Mendelian transgene inheritance pattern. The transformation frequency was 1%, and 6-8 weeks were required for the generation of transgenics.

  3. Genetic transformation of loblolly pine using mature zygotic embryo explants by Agrobacterium tumefaciens

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Agrobacterium tumefaciens strain LBA 4404 carrying pBI121 plasmid was used to transform mature zygotic embryos of three genotypes (E-Hb, E-Ma, and E-Mc) of loblolly pine. The results demonstrated that the expression frequency of b-glucuronidase reporter gene (GUS) varied among genotypes after mature zygotic em-bryos were infected with Agrobacterium tumefaciens cultures. The highest frequency (27.8%) of GUS expressing embryos was obtained from genotype E-Mc with mean number of 21.9 blue GUS spots per embryo. Expression of b-glucuronidase reporter gene was observed on cotyledons, hypocotyls, and radicles of transformed mature zy-gotic embryos, as well as on organogenic callus and regenerated shoots derived from co-cultivated mature zygotic embryos. Nineteen regenerated transgenic plants were obtained from GUS expression and kanamycin resistant calli. The presence and integration of the GUS gene was confirmed by polymerase chain reaction (PCR) and Southern blot analysis. These results suggested that an efficient Agrobacterium tumefaciens-mediated transfor-mation protocol for stable integration of foreign genes into loblolly pine has been developed and that this transfor-mation system could be useful for the future studies on transferring economically important genes to loblolly pine.

  4. Genetic transformation of apple (Malus x domestica) without use of a selectable marker gene

    Science.gov (United States)

    Selectable marker genes are widely used for the efficient transformation of crop plants. In most cases, antibiotic or herbicide resistance marker genes are preferred, because they tend to be most efficient. Due mainly to consumer and grower concerns, considerable effort is being put into developin...

  5. Control of Competence for DNA Transformation in Streptococcus suis by Genetically Transferable Pherotypes

    NARCIS (Netherlands)

    Zaccaria, E.; Baarlen, van P.; Greeff, de A.; Morrison, D.A.; Smith, H.; Wells, J.M.

    2014-01-01

    Here we show that S. suis, a major bacterial pathogen of pigs and emerging pathogen in humans responds to a peptide pheromone by developing competence for DNA transformation. This species does not fall within any of the phylogenetic clusters of streptococci previously shown to regulate competence vi

  6. Establishment of Genetic Transformation System for Miscanthus sacchariflorus and Obtaining of Its Transgenic Plants

    Institute of Scientific and Technical Information of China (English)

    Yi Zili(易自力); Zhou Puhua; Chu Chengcai; Li Xiang; Tian Wenzhong; Wang Li; Cao Shouyun; Tang Zuoshun

    2004-01-01

    The initiation and regeneration system for embryogenic callus of Miscanthus sacchariflrus is established at very high frequency. Potato proteinaseⅡ(pinⅡ) genes are introduced into the callus of Miscanthus sacchariflorus and some transgenic plants are obtained by the particle bombardment transformation system. PCR and Southern analysis show that target genes are integrated into the genome of these transgenic plants.

  7. Advancement of Phenotype Transformation of Cancer-associated Fibroblasts: 
from Genetic Alterations to Epigenetic Modification

    Directory of Open Access Journals (Sweden)

    Dali CHEN

    2015-02-01

    Full Text Available In the field of human cancer research, even though the vast majority attentions were paid to tumor cells as “the seeds”, the roles of tumor microenvironments as “the soil” are gradually explored in recent years. As a dominant compartment of tumor microenvironments, cancer-associated fibroblasts (CAFs were discovered to correlated with tumorigenesis, tumor progression and prognosis. And the exploration of the mechanisms of CAF phenotype transformation would conducive to the further understand of the CAFs function in human cancers. As we known that CAFs have four main origins, including epithelial cells, endothelial cells, mesenchymal stem cells (MSCs and local mesenchymal cells. However, researchers found that all these origins finally conduct similiar phenotypes from intrinsic to extrinsic ones. Thus, what and how a mechanism can conduct the phenotype transformation of CAFs with different origins? Two viewpoints are proposed to try to answer the quetsion, involving genetic alterations and epigenetic modifications. This review will systematically summarize the advancement of mechanisms of CAF phenotype transformations in the aspect of genentic and epigenetic modifications.

  8. Cell transformation and mutability of different genetic loci in mammalian cells by metabolically activated carcinogenic polycylic hydrocarbons

    Energy Technology Data Exchange (ETDEWEB)

    Huberman, E.

    1977-01-01

    Treatment of experimental animals with chemical carcinogens, including some polycyclic hydrocarbons, can result in the formation of malignant tumors. The process whereby some chemicals induce malignancy is as yet unknown. However, in a model system using mammalian cells in culture, it was possible to show that the chemical carcinogens induce malignant transformation rather than select for pre-existing tumor cells. In the process of the in vitro cell transformation, the normal cells, which have an oriented pattern of cell growth, a limited life-span in vitro, and are not tumorigenic, are converted into cells that have a hereditary random pattern of cell growth, the ability to grow continuously in culture, and the ability to form tumors in vivo. This stable heritable phenotype of the transformed cells is similar to that of cells derived from spontaneous or experimentally induced tumors. Such stable heritable phenotype changes may arise from alteration in gene expression due to a somatic mutation after interaction of the carcinogen with cellular DNA. In the present experiments we have shown that metabolically activated carcinogenic polycyclic hydrocarbons which have been shown to bind to cellular DNA induce somatic mutations at different genetic loci in mammalian cells and that there is a relationship between the degree of mutant induction and the degree of carcinogenicity of the different hydrocarbons tested.

  9. Blue Genes: An Integrative Laboratory to Differentiate Genetic Transformation from Gene Mutation for Underclassmen

    Science.gov (United States)

    Militello, Kevin T.; Chang, Ming-Mei; Simon, Robert D.; Lazatin, Justine C.

    2016-01-01

    The ability of students to understand the relationship between genotype and phenotype, and the mechanisms by which genotypes and phenotypes can change is essential for students studying genetics. To this end, we have developed a four-week laboratory called Blue Genes, which is designed to help novice students discriminate between two mechanisms by…

  10. Genetic transformation of selected mature cork oak (Quercus suber L.) trees.

    Science.gov (United States)

    Alvarez, R; Alonso, P; Cortizo, M; Celestino, C; Hernández, I; Toribio, M; Ordás, R J

    2004-10-01

    A transformation system for selected mature cork oak (Quercus suber L.) trees using Agrobacterium tumefaciens has been established. Embryos obtained from recurrent proliferating embryogenic masses were inoculated with A. tumefaciens strains EHA105, LBA4404 or AGL1 harbouring the plasmid pBINUbiGUSint [carrying the neomycin phosphotransferase II (nptII) and beta-glucuronidase (uidA) genes]. The highest transformation efficiency (4%) was obtained when freshly isolated explants were inoculated with A. tumefaciens strain AGL1. Evidence of stable transgene integration was obtained by PCR for the nptII and uidA genes, Southern blotting and expression of the uidA gene. The transgenic embryos were germinated and successfully transferred to soil.

  11. Advances in Agrobacterium tumefaciens-mediated genetic transformation of graminaceous crops.

    Science.gov (United States)

    Singh, Roshan Kumar; Prasad, Manoj

    2016-05-01

    Steady increase in global population poses several challenges to plant science research, including demand for increased crop productivity, grain yield, nutritional quality and improved tolerance to different environmental factors. Transgene-based approaches are promising to address these challenges by transferring potential candidate genes to host organisms through different strategies. Agrobacterium-mediated gene transfer is one such strategy which is well known for enabling efficient gene transfer in both monocot and dicots. Due to its versatility, this technique underwent several advancements including development of improved in vitro plant regeneration system, co-cultivation and selection methods, and use of hyper-virulent strains of Agrobacterium tumefaciens harbouring super-binary vectors. The efficiency of this method has also been enhanced by the use of acetosyringone to induce the activity of vir genes, silver nitrate to reduce the Agrobacterium-induced necrosis and cysteine to avoid callus browning during co-cultivation. In the last two decades, extensive efforts have been invested towards achieving efficient Agrobacterium-mediated transformation in cereals. Though high-efficiency transformation systems have been developed for rice and maize, comparatively lesser progress has been reported in other graminaceous crops. In this context, the present review discusses the progress made in Agrobacterium-mediated transformation system in rice, maize, wheat, barley, sorghum, sugarcane, Brachypodium, millets, bioenergy and forage and turf grasses. In addition, it also provides an overview of the genes that have been recently transferred to these graminaceous crops using Agrobacterium, bottlenecks in this technique and future possibilities for crop improvement.

  12. Studies on genetic transformation of coffee by using electroporation and the bioistic method.

    OpenAIRE

    Boxtel, van, A.J.B.

    1994-01-01

    The present study aimed simultaneously at an improvement of coffee regeneration systems and at a definition of factors influencing the efficiency of direct gene transfer methods. The development of an improved regeneration system, based on high frequency somatic embryogenesis from leaf explants, passing through multiplication of embryogenic callus in liquid medium, is described. This method can contribute to the obtaining of high protoplast yields and offers perspectives for use in genetic tr...

  13. AGROBACTERIUM-MEDIATED GENETIC TRANSFORMATION OF SORGHUM USING TISSUE CULTURE-BASED AND POLLEN-MEDIATED APPROACHES

    Directory of Open Access Journals (Sweden)

    Elkonin L.A.

    2012-08-01

    Full Text Available Genetic transformation is a powerful tool for genetic improvement of arable crops. Genetic engineering approaches are especially important for modification of starch and protein contents, vitamin and micronutrient concentration, improvement of nutritive value of protein fractions, and increase tolerance to environmental stresses. Application of transgenic technologies for genetic improvement of sorghum, a highly productive heat tolerant and drought resistant crop, is extremely important since climate aridization in many regions all over the globe hampers sustainable production of traditional cereals, such as wheat, maize and barley. However, sorghum, in spite of great number of investigations, is one of the most recalcitrant crop species to genetic modification. The most frequently reported problems are a low frequency of transformation and silencing of transgenes. Using the A. tumefaciens strain AGL0/p35SGIB with the bar and gus-intron genes under the nos and CaMV35S promoters, respectively, we studied different methods of Agrobacterium-mediated genetic transformation of the grain sorghum: in vitro culture-based techniques, by inoculation of immature embryos or embryo-derived calli, and pollen-mediated approach, by inoculation of flowering panicles. Four lines of grain sorghum – Milo-10, [9E] Milo-10 (CMS-line, KVV-114, and KVV-45 – were used. In both approaches, for activation of vir-genes agrobacterial cell suspension was grown in the AB or modified AB media with acetosyringone at room temperature. In vitro culture approach was effective for obtaining transgenic plants in the lines Milo-10 and KVV-45, which were able to produce embryogenic callus from immature embryos after their co-cultivation with agrobacterial cell suspension. Callus cultures tolerant to glufosinate ammonium (GA and capable to plant regeneration were obtained. The frequency of immature embryos producing PCR-positive transgenic plants varied in different experiments

  14. Transformação genética de cereais via Agrobacterium tumefaciens Cereal genetic transformation via Agrobacterium tumefaciens

    Directory of Open Access Journals (Sweden)

    Cristine Luise Handel

    1997-06-01

    Full Text Available A transformação genética via Agrobacterium tumefaciens é um método que permite a inserção de uma ou poucas cópias do transgene no DNA da planta hospedeira. Esta pode ser uma ferramenta importante para os melhoristas, pois, além de aumentar a variabilidade genética existente, torna possível criar variabilidade não disponível via métodos de melhoramento convencional. No entanto, ainda existem algumas dificuldades a serem superadas para que os genes de interesse agronômico sejam incorporados no genoma dos cereais, como aidentificação de estirpes de bactérias que infectem monocotiledôneas e a adequação da técnica. O objetivo deste trabalho é de revisar as potencialidades e problemas do uso da A. tumefaciens para transformação de cereais no presentemomento e abordar suas perspectivas futuras. Trabalhos recentes com arroz e trigo indicam que estas culturas podem ser transformadas com A. tumefaciens, sendo que em arroz plantas transgênicas foram obtidas com este método. Esta tecnologia vem sendo aprimorada e a curto prazo possibilitará a transferência de genes para diversas espécies monocotiledôneas.The genetic transformation via Agrobacterium tumefaciens allows the insertion of one or few copies of a transgene into the host DNA. This can be an important tool to plant breeders because it expands the genetic variability in breeding programs, developing variability not readily available from traditional methods. However, there are some difficulties that have to be overcome before this technology may be used in cereals, as the identification of highly ineffective bacteria strains and the adjustments of the technique to various crops. The objective of this paper is to revise the potentialities and limitations of using A. tumefaciens to transform cereals and to indicate the future perspectives of this technology. Recent studies have indicated that it is possible to transform rice and wheat with A. tumefaciens, so that rice

  15. Appearance and characterization of fruit image textures for quality sorting using wavelet transform and genetic algorithms.

    Science.gov (United States)

    Khoje, Suchitra

    2017-07-24

    Images of four qualities of mangoes and guavas are evaluated for color and textural features to characterize and classify them, and to model the fruit appearance grading. The paper discusses three approaches to identify most discriminating texture features of both the fruits. In the first approach, fruit's color and texture features are selected using Mahalanobis distance. A total of 20 color features and 40 textural features are extracted for analysis. Using Mahalanobis distance and feature intercorrelation analyses, one best color feature (mean of a* [L*a*b* color space]) and two textural features (energy a*, contrast of H*) are selected as features for Guava while two best color features (R std, H std) and one textural features (energy b*) are selected as features for mangoes with the highest discriminate power. The second approach studies some common wavelet families for searching the best classification model for fruit quality grading. The wavelet features extracted from five basic mother wavelets (db, bior, rbior, Coif, Sym) are explored to characterize fruits texture appearance. In third approach, genetic algorithm is used to select only those color and wavelet texture features that are relevant to the separation of the class, from a large universe of features. The study shows that image color and texture features which were identified using a genetic algorithm can distinguish between various qualities classes of fruits. The experimental results showed that support vector machine classifier is elected for Guava grading with an accuracy of 97.61% and artificial neural network is elected from Mango grading with an accuracy of 95.65%. The proposed method is nondestructive fruit quality assessment method. The experimental results has proven that Genetic algorithm along with wavelet textures feature has potential to discriminate fruit quality. Finally, it can be concluded that discussed method is an accurate, reliable, and objective tool to determine fruit

  16. Transformation of personal computers and mobile phones into genetic diagnostic systems.

    Science.gov (United States)

    Walker, Faye M; Ahmad, Kareem M; Eisenstein, Michael; Soh, H Tom

    2014-09-16

    Molecular diagnostics based on the polymerase chain reaction (PCR) offer rapid and sensitive means for detecting infectious disease, but prohibitive costs have impeded their use in resource-limited settings where such diseases are endemic. In this work, we report an innovative method for transforming a desktop computer and a mobile camera phone--devices that have become readily accessible in developing countries--into a highly sensitive DNA detection system. This transformation was achieved by converting a desktop computer into a de facto thermal cycler with software that controls the temperature of the central processing unit (CPU), allowing for highly efficient PCR. Next, we reconfigured the mobile phone into a fluorescence imager by adding a low-cost filter, which enabled us to quantitatively measure the resulting PCR amplicons. Our system is highly sensitive, achieving quantitative detection of as little as 9.6 attograms of target DNA, and we show that its performance is comparable to advanced laboratory instruments at approximately 1/500th of the cost. Finally, in order to demonstrate clinical utility, we have used our platform for the successful detection of genomic DNA from the parasite that causes Chagas disease, Trypanosoma cruzi, directly in whole, unprocessed human blood at concentrations 4-fold below the clinical titer of the parasite.

  17. Genetic identification of arsenate reductase and arsenite oxidase in redox transformations carried out by arsenic metabolising prokaryotes - A comprehensive review.

    Science.gov (United States)

    Kumari, Nisha; Jagadevan, Sheeja

    2016-11-01

    Arsenic (As) contamination in water is a cause of major concern to human population worldwide, especially in Bangladesh and West Bengal, India. Arsenite (As(III)) and arsenate (As(V)) are the two common forms in which arsenic exists in soil and groundwater, the former being more mobile and toxic. A large number of arsenic metabolising microorganisms play a crucial role in microbial transformation of arsenic between its different states, thus playing a key role in remediation of arsenic contaminated water. This review focuses on advances in biochemical, molecular and genomic developments in the field of arsenic metabolising bacteria - covering recent developments in the understanding of structure of arsenate reductase and arsenite oxidase enzymes, their gene and operon structures and their mechanism of action. The genetic and molecular studies of these microbes and their proteins may lead to evolution of successful strategies for effective implementation of bioremediation programs. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Genetics

    Science.gov (United States)

    ... Inheritance; Heterozygous; Inheritance patterns; Heredity and disease; Heritable; Genetic markers ... The chromosomes are made up of strands of genetic information called DNA. Each chromosome contains sections of ...

  19. Genetic Transformation of Watermelon with Pumpkin DNA by Low Energy Ion Beam-mediated Introduction

    Institute of Scientific and Technical Information of China (English)

    王浩波; 高秀武; 郭金华; 黄群策; 余增亮

    2002-01-01

    The No.601 watermelon (citrullus lanatus) seeds were treated with 25 keV N+ implantation at the dosage of 7.8 × 1016 ions/cm2. After treatment, watermelon seeds were incubated with 380 μg/μl pumpkin (Cucubita, maxima Duch) DNA solution at 35 ℃ for 5 hours. By twogenerations of selection and resistance screening at seedling stage, one transformed material was selected out, whose rind color is similar to that of the donor pumpkin and whose size of seeds is between that of the donor and the receptor. Using AFLP (amplified fragment length polymorphism) technique, two polymorphic DNA fragments were amplified. This primarily testified that the donor DNA fragments/gene were introduced into the receptor cell and integrated into the genomic DNA of the receptor.

  20. Genetic Transformation of Watermelon with Pumpkin DNA by Low Energy Ion Beam-Mediated Introduction

    Science.gov (United States)

    Wang, Hao-bo; Gao, Xiu-wu; Guo, Jin-hua; Huang, Qun-ce; Yu, Zeng-liang

    2002-12-01

    The No.601 watermelon (citrullus lanatus) seeds were treated with 25 keV N+ implantation at the dosage of 7.8 × 1016 ions/cm2. After treatment, watermelon seeds were incubated with 380 μg/μl pumpkin (Cucubita, maxima Duch) DNA solution at 35 °C for 5 hours. By two-generations of selection and resistance screening at seedling stage, one transformed material was selected out, whose rind color is similar to that of the donor pumpkin and whose size of seeds is between that of the donor and the receptor. Using AFLP (amplified fragment length polymorphism) technique, two polymorphic DNA fragments were amplified. This primarily testified that the donor DNA fragments/gene were introduced into the receptor cell and integrated into the genomic DNA of the receptor.

  1. Streptococcus pneumoniae R6 interspecies transformation: genetic analysis of penicillin resistance determinants and genome-wide recombination events.

    Science.gov (United States)

    Sauerbier, Julia; Maurer, Patrick; Rieger, Martin; Hakenbeck, Regine

    2012-11-01

    Interspecies gene transfer has been implicated as the major driving force for the evolution of penicillin resistance in Streptococcus pneumoniae. Genomic alterations of S. pneumoniae R6 introduced during four successive transformations with DNA of the high-level penicillin-resistant Streptococcus mitis B6 with beta-lactam selection have now been determined and the contribution of genes to high resistance levels was analysed genetically. Essential for high level resistance to penicillins of the transformant CCCB was the combination of murM(B) (6) and the 3' region of pbp2b(B) (6) . Sequences of both genes were detected in clinical isolates of S. pneumoniae, confirming the participation of S. mitis in the global gene pool of beta-lactam resistance determinants. The S. mitis PBP1b gene which contains an authentic stop codon within the transpeptidase domain is now shown to contribute only marginal to resistance, but it is possible that the presence of its transglycosylase domain is important in the context of cognate PBPs. The genome sequence of CCCB revealed 36 recombination events, including deletion and acquisition of genes and repeat elements. A total of 78 genes were affected representing 67 kb or 3.3% of the genome, documenting extensive alterations scattered throughout the genome.

  2. A Cytotoxic and Anti-inflammatory Campesterol Derivative from Genetically Transformed Hairy Roots of Lopezia racemosa Cav. (Onagraceae

    Directory of Open Access Journals (Sweden)

    Norma Elizabeth Moreno-Anzúrez

    2017-01-01

    Full Text Available The genetically transformed hairy root line LRT 7.31 obtained by infecting leaf explants of Lopezia racemosa Cav with the Agrobacterium rhizogenes strain ATCC15834/pTDT, was evaluated to identify the anti-inflammatory and cytotoxic compounds reported previously for the wild plant. After several subcultures of the LRT 7.31 line, the bio-guided fractionation of the dichloromethane–methanol (1:1 extract obtained from dry biomass afforded a fraction that showed important in vivo anti-inflammatory, and in vitro cytotoxic activities. Chemical separation of the active fraction allowed us to identify the triterpenes ursolic (1 and oleanolic (2 acids, and (23R-2α,3β,23,28-tetrahydroxy-14,15-dehydrocampesterol (3 as the anti-inflammatory principles of the active fraction. A new molecule 3 was characterized by spectroscopic analysis of its tetraacetate derivative 3a. This compound was not described in previous reports of callus cultures, in vitro germinated seedlings and wild plant extracts of whole L. racemosa plants. The anti-inflammatory and cytotoxic activities displayed by the fraction are associated to the presence of compounds 1–3. The present study reports the obtaining of the transformed hairy roots, the bioguided isolation of the new molecule 3, and its structure characterization.

  3. Ultraviolet sensitivity of the addition, deletion and replacement of long nonhomologous DNA segments by genetic transformation of Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Walter, R.B.; Stuy, J.H. (Florida State Univ., Tallahassee (USA). Dept. of Biological Science)

    1982-03-01

    The construction and some properties of Haemophilus influenzae Rd strains with long and different R plasmid-derived DNA segments (nonhomologous inserts) at the same site in the HP1 prophage have previously been described. These inserts can be added to a recipient's genome by genetic transformation, they can be deleted from the recipient genome, or they can be replaced by another insert. It is reported that the UV inactivation of all three phenomena followed single hit kinetics. Deletion was roughly 10 times more resistant; its UV-sensitivity equalled that of a high-efficiency point mutation. There was an inverse correlation between UV-sensitivity and additive transformation efficiency of the various inserts; sensitivity may thus be a measure of insert size. This correlation was not seen for deletion. All three phenomena were more sensitive when they were measured on excision repair-deficient uvr/sup -/ recipients. The dose-reduction factor for addition was about 1.5 while it was about 2.6 for deletion.

  4. Transformation, evaluation of GTGene and multivariate genetic analysis for morpho-physiological and yield attributing traits in Zea mays

    Directory of Open Access Journals (Sweden)

    Aaliya Khadija

    2016-01-01

    Full Text Available Maize is the 3rd major crop grown all over the world that fulfills the needs of millions of people. Various biotic and abiotic factors caused reduction in grain yield of maize, among them weeds show most adverse effects. Objective of this study was to develop glyphosate resistance maize for developing maize hybrids and synthetic varieties with high grain and fodder yield potential. Glyphosate is a broad spectrum herbicide and resistance of crop against the herbicide, allows post emergence application, which otherwise is impossible. Higher GTG crude protein (479.94μg/g was recorded for CIL 194-975 plant. The plant CIL 194-1786 showed higher expression level of GTG protein, indicated that transformation efficiency was better for line CIL-194 than CIL-123. GTGene stably integrated to the genome of the maize lines CIL-194 and CIL-123, these transformed lines showed more ability to cope with herbicides as compared with non-transformed lines in greenhouse and field trials. The transgenic lines were sown in field and data was recorded for various morpho-physiological, grain, fodder yield and quality traits. Data was statistically analyzed to check significance of results at <0.05% probability level. It was concluded from heritability, genetic advance, stepwise multiple linear regression, principle component and factor analysis that the selection on the basis of grain yield, green fodder yield, stem diameter, cob weight, stomata conductance, chlorophyll contents, photosynthetic rate, cob length, grain protein, grain oil, embryo percentage and grain starch may be fruitful keeping in view as the major contributing traits to improve crop yield and production. It was suggested that the transgenic lines for glyphosate resistance may be used to develop transgenic hybrids and synthetic varieties with higher grain yield with high quality.

  5. Partial discharge localization in power transformers based on the sequential quadratic programming-genetic algorithm adopting acoustic emission techniques

    Science.gov (United States)

    Liu, Hua-Long; Liu, Hua-Dong

    2014-10-01

    Partial discharge (PD) in power transformers is one of the prime reasons resulting in insulation degradation and power faults. Hence, it is of great importance to study the techniques of the detection and localization of PD in theory and practice. The detection and localization of PD employing acoustic emission (AE) techniques, as a kind of non-destructive testing, plus due to the advantages of powerful capability of locating and high precision, have been paid more and more attention. The localization algorithm is the key factor to decide the localization accuracy in AE localization of PD. Many kinds of localization algorithms exist for the PD source localization adopting AE techniques including intelligent and non-intelligent algorithms. However, the existed algorithms possess some defects such as the premature convergence phenomenon, poor local optimization ability and unsuitability for the field applications. To overcome the poor local optimization ability and easily caused premature convergence phenomenon of the fundamental genetic algorithm (GA), a new kind of improved GA is proposed, namely the sequence quadratic programming-genetic algorithm (SQP-GA). For the hybrid optimization algorithm, SQP-GA, the sequence quadratic programming (SQP) algorithm which is used as a basic operator is integrated into the fundamental GA, so the local searching ability of the fundamental GA is improved effectively and the premature convergence phenomenon is overcome. Experimental results of the numerical simulations of benchmark functions show that the hybrid optimization algorithm, SQP-GA, is better than the fundamental GA in the convergence speed and optimization precision, and the proposed algorithm in this paper has outstanding optimization effect. At the same time, the presented SQP-GA in the paper is applied to solve the ultrasonic localization problem of PD in transformers, then the ultrasonic localization method of PD in transformers based on the SQP-GA is proposed. And

  6. Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of bacopa saponins in transformed calli and plants.

    Science.gov (United States)

    Majumdar, Sukanya; Garai, Saraswati; Jha, Sumita

    2011-05-01

    We have developed an efficient transformation system for Bacopa monnieri, an important Indian medicinal plant, using Agrobacterium rhizogenes strains LBA 9402 and A4. Transformed roots induced by strain LBA 9402 spontaneously dedifferentiated to callus while excised roots induced by strain A4 spontaneously showed induction of shoot buds within 10 days. PCR and RT-PCR analysis revealed the presence and expression of the rolAB and rolC genes at the transcription level in pRi A4 transformed cultures indicating that the TL-DNA was integrated retained and expressed in the A4-Ri transformed shoots. Transformed calli showed the presence of rolAB or rol A, TR and ags genes. Transformed plants showed morphological features typically seen in transgenic plants produced by A. rhizogenes. Growth and biomass accumulation was significantly higher in the transformed shoots (twofold) and roots (fourfold) than in the non-transformed (WT) plants. In pRi A4-transformed plants, the content of bacopasaponin D, bacopasaponin F, bacopaside II and bacopaside V was enhanced significantly as compared to WT plants of similar age while bacoside A3 and bacopasaponin C content was comparable with that of WT plants. Significant increase in content of five bacopa saponins could be detected in pRi 9402-transformed callus cultures. There is an overall stimulatory effect on accumulation of bacopa saponins in transformed plants and cells of B. monnieri establishing the role of endogenous elicitation by Ri T-DNA of A. rhizogenes.

  7. Unlocking the potential of tropical root crop biotechnology in east Africa by establishing a genetic transformation platform for local farmer-preferred cassava cultivars.

    Science.gov (United States)

    Nyaboga, Evans; Njiru, Joshua; Nguu, Edward; Gruissem, Wilhelm; Vanderschuren, Herve; Tripathi, Leena

    2013-01-01

    Cassava genetic transformation capacity is still mostly restricted to advanced laboratories in the USA, Europe and China; and its implementation and maintenance in African laboratories has remained scarce. The impact of transgenic technologies for genetic improvement of cassava will depend largely on the transfer of such capabilities to researchers in Africa, where cassava has an important socioeconomic niche. A major constraint to the development of genetic transformation technologies for cassava improvement has been the lack of an efficient and robust transformation and regeneration system. Despite the success achieved in genetic modification of few cassava cultivars, including the model cultivar 60444, transgenic cassava production remains difficult for farmer-preferred cultivars. In this study, a protocol for cultivar 60444 developed at ETH Zurich was successfully implemented and optimized to establish transformation of farmer-preferred cassava cultivars popular in east Africa. The conditions for production and proliferation of friable embryogenic calli (FEC) and Agrobacterium-mediated transformation were optimized for three east African farmer-preferred cultivars (Ebwanatereka, Kibandameno and Serere). Our results demonstrated transformation efficiencies of about 14-22 independent transgenic lines per 100 mg of FEC for farmer-preferred cultivars in comparison to 28 lines per 100 mg of the model cultivar 60444. The presence, integration and expression of the transgenes were confirmed by PCR, Southern blot analysis and histochemical GUS assay. This study reports the establishment of a cassava transformation platform at International Institute of Tropical Agriculture (IITA) hosted by Biosciences eastern and central Africa (BecA) hub in Kenya and provides the basis for transferring important traits such as virus resistance and prolonged shelf-life to farmer-preferred cultivars in east Africa. We anticipate that such platform will also be instrumental to transfer

  8. Unlocking the potential of tropical root crop biotechnology in east Africa by establishing a genetic transformation platform for local farmer-preferred cassava cultivars

    Directory of Open Access Journals (Sweden)

    Evans eNyaboga

    2013-12-01

    Full Text Available Cassava genetic transformation capacity is still mostly restricted to advanced laboratories in the USA, Europe and China; and its implementation and maintainance in African laboratories has remained scarce. The impact of transgenic technologies for genetic improvement of cassava will depend largely on the transfer of such capabilities to researchers in Africa, where cassava has an important socioeconomic niche. A major constraint to the development of genetic transformation technologies for cassava improvement has been the lack of an efficient and robust transformation and regeneration system. Despite the success achieved in genetic modification of few cassava cultivars, including the model cultivar 60444, transgenic cassava production remains difficult for farmer-preferred cultivars. In this study, a protocol for cultivar 60444 developed at ETH Zurich was successfully implemented and optimized to establish transformation of farmer-preferred cassava cultivars popular in east Africa. The conditions for production and proliferation of friable embryogenic calli (FEC and Agrobacterium-mediated transformation were optimized for three east African farmer-preferred cultivars (Ebwanatereka, Kibandameno and Serere. Our results demonstrated transformation efficiencies of about 14-22 independent transgenic lines per 100 mg of FEC for farmer-preferred cultivars in comparison to 28 lines per 100 mg of the model cultivar 60444. The presence, integration and expression of the transgenes were confirmed by PCR, Southern blot analysis and histochemical GUS assay. This study reports the establishment of a cassava transformation platform at International Institute of Tropical Agriculture (IITA hosted by Biosciences eastern and central Africa (BecA hub in Kenya and provides the basis for transferring important traits such as virus resistance and prolonged shelf-life to farmer-preferred cultivars in east Africa. We anticipate that such platform will also be

  9. Genetic transformation and analysis of rice OsAPx2 gene in Medicago sativa.

    Directory of Open Access Journals (Sweden)

    Qingjie Guan

    Full Text Available The OsAPx2 gene from rice was cloned to produce PBI121::OsAPx2 dual-expression plants, of which expression level would be increasing under stressful conditions. The enzyme ascorbate peroxidase (APX in the leaves and roots of the plants increased with increasing exposure time to different sodium chloride (NaCl and hydrogen peroxide (H(2O(2concentrations, as indicated by protein gel blot analysis. The increased enzyme yield improved the ability of the plants to resist the stress treatments. The OsAPx2 gene was localized in the cytoplasm of epidermal onion cells as indicated by the instantaneous expression of green fluorescence. An 80% regeneration rate was observed in Medicago sativa L. plants transformed with the OsAPx2 gene using Agrobacterium tumefaciens, as indicated by specific primer PCR. The OsAPx2 gene was expressed at the mRNA level and the individual M. sativa (T#1,T#2,T#5 were obtained through assaying the generation of positive T2 using RNA gel blot analysis. When the seeds of the wild type (WT and the T2 (T#1,T#5 were incubated in culture containing MS with NaCl for 7 days, the results as shown of following: the root length of transgenic plant was longer than WT plants, the H(2O(2 content in roots of WT was more than of transgenic plants, the APX activity under stresses increased by 2.89 times compared with the WT, the malondialdehyde (MDA content of the WT was higher than the transgenic plants, the leaves of the WT turned yellow, but those of the transgenic plants remained green and remained healthy. The chlorophyll content in the WT leaves was less than in the transgenic plants, after soaking in solutions of H(2O(2, sodium sulfite (Na(2SO(3, and sodium bicarbonate (NaHCO(3. Therefore, the OsAPx2 gene overexpression in transgenic M. sativa improves the removal of H(2O(2 and the salt-resistance compared with WT plants. A novel strain of M. sativa carrying a salt-resistance gene was obtained.

  10. Genetic transformation and analysis of rice OsAPx2 gene in Medicago sativa.

    Science.gov (United States)

    Guan, Qingjie; Takano, Tetsuo; Liu, Shenkui

    2012-01-01

    The OsAPx2 gene from rice was cloned to produce PBI121::OsAPx2 dual-expression plants, of which expression level would be increasing under stressful conditions. The enzyme ascorbate peroxidase (APX) in the leaves and roots of the plants increased with increasing exposure time to different sodium chloride (NaCl) and hydrogen peroxide (H(2)O(2))concentrations, as indicated by protein gel blot analysis. The increased enzyme yield improved the ability of the plants to resist the stress treatments. The OsAPx2 gene was localized in the cytoplasm of epidermal onion cells as indicated by the instantaneous expression of green fluorescence. An 80% regeneration rate was observed in Medicago sativa L. plants transformed with the OsAPx2 gene using Agrobacterium tumefaciens, as indicated by specific primer PCR. The OsAPx2 gene was expressed at the mRNA level and the individual M. sativa (T#1,T#2,T#5) were obtained through assaying the generation of positive T2 using RNA gel blot analysis. When the seeds of the wild type (WT) and the T2 (T#1,T#5) were incubated in culture containing MS with NaCl for 7 days, the results as shown of following: the root length of transgenic plant was longer than WT plants, the H(2)O(2) content in roots of WT was more than of transgenic plants, the APX activity under stresses increased by 2.89 times compared with the WT, the malondialdehyde (MDA) content of the WT was higher than the transgenic plants, the leaves of the WT turned yellow, but those of the transgenic plants remained green and remained healthy. The chlorophyll content in the WT leaves was less than in the transgenic plants, after soaking in solutions of H(2)O(2), sodium sulfite (Na(2)SO(3)), and sodium bicarbonate (NaHCO(3)). Therefore, the OsAPx2 gene overexpression in transgenic M. sativa improves the removal of H(2)O(2) and the salt-resistance compared with WT plants. A novel strain of M. sativa carrying a salt-resistance gene was obtained.

  11. Agrobacterium-mediated genetic transformation and regeneration of transgenic plants using leaf midribs as explants in ramie [Boehmeria nivea (L.) Gaud].

    Science.gov (United States)

    An, Xia; Wang, Bo; Liu, Lijun; Jiang, Hui; Chen, Jie; Ye, Shengtuo; Chen, Leiyu; Guo, Pingan; Huang, Xing; Peng, Dingxiang

    2014-05-01

    In this study, leaf midribs, the elite explants, were used for the first time to develop an efficient regeneration and transformation protocol for ramie [Boehmeria nivea (L.) Gaud.] via Agrobacterium-mediated genetic transformation. Sensitivity of leaf midribs regeneration to kanamycin was evaluated, which showed that 40 mg l(-1) was the optimal concentration needed to create the necessary selection pressure. Factors affecting the ramie transformation efficiency were evaluated, including leaf age, Agrobacterium concentration, length of infection time for the Agrobacterium solution, acetosyringone concentration in the co-cultivation medium, and the co-cultivation period. The midrib explants from 40-day-old in vitro shoots, an Agrobacterium concentration at OD600 of 0.6, 10-min immersion in the bacteria solution, an acetosyringone concentration of 50 mg l(-1) in the co-cultivation medium and a 3-day co-cultivation period produced the highest efficiencies of regeneration and transformation. In this study, the average transformation rate was 23.25%. Polymerase chain reactions using GUS and NPTII gene-specific primers, Southern blot and histochemical GUS staining analyses further confirmed that the transgene was integrated into the ramie genome and expressed in the transgenic ramie. The establishment of this system of Agrobacterium-mediated genetic transformation and regeneration of transgenic plants will be used not only to introduce genes of interest into the ramie genome for the purpose of trait improvement, but also as a common means of testing gene function by enhancing or inhibiting the expression of target genes.

  12. Induction of endogenous telomerase (hTERT) by c-Myc in WI-38 fibroblasts transformed with specific genetic elements.

    Science.gov (United States)

    Casillas, Mark A; Brotherton, Scott L; Andrews, Lucy G; Ruppert, J Michael; Tollefsbol, Trygve O

    2003-10-16

    Elucidation of the mechanisms governing expression of the human telomerase reverse transcriptase (hTERT) is important for understanding cancer pathogenesis. Approximately 90% of tumors express hTERT, the major catalytic component of telomerase. Activation of telomerase is an early event, and high levels of this activity correlate with poor prognosis. Recent studies have shown that the transcription factors c-Myc and Mad1 activate and repress hTERT, respectively. It is not clear how these transcription factors compete for the same recognition sequence in the hTERT core promoter region. Studies have shown that the combined expression of SV40 large T antigen (T-Ag), hTERT, and H-Ras is able to transform human cells. In this study, we used a distinct human cell type, WI-38 fetal lung fibroblasts used extensively for senescence studies. We transduced cells with amphotropic retroviral constructs containing SV40 T antigen, hTERT, and activated H-ras. Transduced cells exhibited anchorage independence in soft agar and expressed increased levels of c-Myc and endogenous hTERT. These effects were observed by 25 population doublings (PDs) following the establishment of the neoplastic cell line. During the process of transformation, we observed a switch from Mad1/Max to c-Myc/Max binding to oligonucleotide sequences containing the hTERT promoter distal and proximal E-boxes. c-Myc can bind specifically to the hTERT promoter in vitro, indicating that c-Myc expression in tumors may account for the increased expression of hTERT observed in vivo. These findings indicate that the widely used model system of WI-38 fibroblasts can be employed for transformation studies using defined genetic elements and that the endogenous hTERT and c-Myc are induced in these cells during early tumorigenesis. Such studies should have important implications in the mechanisms of hTERT and c-Myc induction in the beginning stages of tumorigenesis and facilitate extension of these studies to novel models of

  13. Genomic simulation for the study of data transformation efficiency in the genetic evaluation of cattle in the presence of heterogeneity - 10.4025/actascianimsci.v34i1.10664

    Directory of Open Access Journals (Sweden)

    Sebastião Martins Filho

    2011-11-01

    Full Text Available Efficiency of data transformation strategies is evaluated to correct heterogeneity among cattle in the genetic evaluation of bulls, cows and progenies. Four data structures of bovine weaning weights were simulated: herds with and without heterogeneity for means and variances, and herds with and without genetic connectedness. In the genetic evaluations, data were used in the original scale and transformed (Logarithmic, Square root, standardization and ratio by phenotypic standard deviation. The evaluated data transformation strategies were not efficient in eliminating the negative effects of heterogeneity among cattle in the genetic evaluation of bulls, cows and progenies.

  14. Improved phytoaccumulation of cadmium by genetically modified tobacco plants (Nicotiana tabacum L.). Physiological and biochemical response of the transformants to cadmium toxicity

    Energy Technology Data Exchange (ETDEWEB)

    Gorinova, N. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria)]. E-mail: noraig60@yahoo.co.uk; Nedkovska, M. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Todorovska, E. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Simova-Stoilova, L. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Stoyanova, Z. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Georgieva, K. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Demirevska-Kepova, K. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Atanassov, A. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Herzig, R. [Phytotech-Foundation PT-F, Quartiergasse 12, CH 3013 Bern (Switzerland)

    2007-01-15

    The response of tobacco plants (Nicotiana tabacum L.)-non-transformed and transformed with a metallothionein gene MThis from Silene vulgaris L. - to increase cadmium supply in the nutrient solution was compared. The transgenic plants accumulated significantly more Cd both in the roots and the leaves. Visual toxicity symptoms and disturbance in water balance were correlated with Cd tissue content. Treatment with 300 {mu}M CdCl{sub 2} resulted in inhibition of photosynthesis and mobilization of the ascorbate-glutathione cycle. Treatment with 500 {mu}M CdCl{sub 2} led to irreversible damage of photosynthesis and oxidative stress. An appearance of a new peroxidase isoform and changes in the leaf polypeptide pattern were observed at the highest Cd concentration. The level of non-protein thiols gradually increased following the Cd treatment both in transgenic and non-transformed plants. - Genetic transformation of Nicotiana tabacum L. by metallothionein gene improved phytoaccumulation of cadmium.

  15. A tyrosine-rich cell surface protein in the diatom Amphora coffeaeformis identified through transcriptome analysis and genetic transformation.

    Directory of Open Access Journals (Sweden)

    Matthias T Buhmann

    Full Text Available Diatoms are single-celled eukaryotic microalgae that are ubiquitously found in almost all aquatic ecosystems, and are characterized by their intricately structured SiO2 (silica-based cell walls. Diatoms with a benthic life style are capable of attaching to any natural or man-made submerged surface, thus contributing substantially to both microbial biofilm communities and economic losses through biofouling. Surface attachment of diatoms is mediated by a carbohydrate- and protein- based glue, yet no protein involved in diatom underwater adhesion has been identified so far. In the present work, we have generated a normalized transcriptome database from the model adhesion diatom Amphora coffeaeformis. Using an unconventional bioinformatics analysis we have identified five proteins that exhibit unique amino acid sequences resembling the amino acid composition of the tyrosine-rich adhesion proteins from mussel footpads. Establishing the first method for the molecular genetic transformation of A. coffeaeformis has enabled investigations into the function of one of these proteins, AC3362, through expression as YFP fusion protein. Biochemical analysis and imaging by fluorescence microscopy revealed that AC3362 is not involved in adhesion, but rather plays a role in biosynthesis and/or structural stability of the cell wall. The methods established in the present study have paved the way for further molecular studies on the mechanisms of underwater adhesion and biological silica formation in the diatom A. coffeaeformis.

  16. Genetic transformation of Fusarium avenaceum by Agrobacterium tumefaciens mediated transformation and the development of a USER-Brick vector construction system

    DEFF Research Database (Denmark)

    Sorensen, Lisette Quaade; Lysøe, Erik; Larsen, Jesper Erup

    2014-01-01

    single step vector construction strategy relying on Uracil Specific Excision Reagent (USER) Fusion cloning, is developed. Results: The new vector construction system, termed USER-Brick, is based on a limited number of PCR amplified vector fragments (core USER-Bricks) which are combined with PCR generated...... FaPKS3 vector was used for optimizing A. tumefaciens mediated transformation (ATMT) of F. avenaceum with respect to six variables. Acetosyringone concentration, co-culturing time, co-culturing temperature and fungal inoculum were found to significantly impact the transformation frequency. Following...... optimization, an average of 140 transformants per 10(6) macroconidia was obtained in experiments aimed at introducing targeted genome modifications. Targeted deletion of FaPKS6 (FA08709.2) in F. avenaceum showed that this gene is essential for biosynthesis of the polyketide/nonribosomal compound fusaristatin A...

  17. Genetic transformation and expression of transgenic lines of Populus x euramericana with insect-resistance and salt-tolerance genes.

    Science.gov (United States)

    Yang, R L; Wang, A X; Zhang, J; Dong, Y; Yang, M S; Wang, J M

    2016-04-29

    We characterized new transgenic varieties of poplar with multiple insect-resistant and salt stress tolerant genes. Two insect-resistant Bacillus thuringiensis (Bt) genes, Cry1Ac and Cry3A, and a salt-tolerant gene, Betaine aldehyde dehydrogenase (BADH) were inserted into a vector, p209-Cry1Ac-Cry3A-BADH. The clone of Populus x euramericana was transformed by the vector using the Agrobacterium-mediated method. Three transgenic lines were assessed using genetic detection and resistance expression analysis. PCR revealed that exogenous genes Cry1Ac, Cry3A, BADH and selective marker gene NPTII were present in three transgenic lines. Quantitative real-time PCR (qPCR) showed significant differences in the transcriptional abundance of three exogenous genes in different lines. Results of assays for Bt toxic proteins showed that the Cry1Ac and Cry3A toxic protein content of each line was 12.83-26.32 and 2108.91-2724.79 ng/g, respectively. The Cry1Ac toxic protein content of different lines was significantly different; the Cry3A toxic protein content was about 100 times higher than that of the Cry1Ac toxic protein. The insect-resistance test revealed the mortality rate of transgenic lines to Hyphantria cunea L1 larvae varied by 42.2-66.7%, which was significantly higher than non-transgenic lines. The mortality rate of L1 and L2 Plagiodera versicolora larvae was 100%. The insecticidal effect of transgenic lines to P. versicolora larvae was higher than that to H. cunea larvae. NaCl stress tolerance of three transgenic lines under 3-6% NaCl concentration was significantly higher than that of non-transgenic lines.

  18. An in vivo Transient Expression System Can Be Applied for Rapid and Effective Selection of Artificial MicroRNA Constructs for Plant Stable Genetic Transformation

    Institute of Scientific and Technical Information of China (English)

    Basdeo Bhagwat; Ming Chi; Li Su; Haifeng Tang; Guiliang Tang; Yu Xiang

    2013-01-01

    The utility of artificial microRNAs (amiRNAs) to induce loss of gene function has been reported for many plant species,but expression efficiency of the different amiRNA constructs in different transgenic plants was less predictable.In this study,expressions of amiRNAs through the gene backbone of Arabidopsis miR 168a were examined by both Agrobacterium-mediated transient expression and stable plant genetic transformation.A corresponding trend in expression of amiRNAs by the same amiRNA constructs between the transient and the stable expression systems was observed in the experiments.Plant genetic transformation of the constructs that were highly expressible in amiRNAs in the transient agro-infiltration assays resulted in generation of transgenic lines with high level of amiRNAs.This provides a simple method for rapid and effective selection of amiRNA constructs used for a time-consuming genetic transformation in plants.

  19. [Hyper spectral estimation method for soil alkali hydrolysable nitrogen content based on discrete wavelet transform and genetic algorithm in combining with partial least squares DWT-GA-PLS)].

    Science.gov (United States)

    Chen, Hong-Yan; Zhao, Geng-Xing; Li, Xi-Can; Wang, Xiang-Feng; Li, Yu-Ling

    2013-11-01

    Taking the Qihe County in Shandong Province of East China as the study area, soil samples were collected from the field, and based on the hyperspectral reflectance measurement of the soil samples and the transformation with the first deviation, the spectra were denoised and compressed by discrete wavelet transform (DWT), the variables for the soil alkali hydrolysable nitrogen quantitative estimation models were selected by genetic algorithms (GA), and the estimation models for the soil alkali hydrolysable nitrogen content were built by using partial least squares (PLS) regression. The discrete wavelet transform and genetic algorithm in combining with partial least squares (DWT-GA-PLS) could not only compress the spectrum variables and reduce the model variables, but also improve the quantitative estimation accuracy of soil alkali hydrolysable nitrogen content. Based on the 1-2 levels low frequency coefficients of discrete wavelet transform, and under the condition of large scale decrement of spectrum variables, the calibration models could achieve the higher or the same prediction accuracy as the soil full spectra. The model based on the second level low frequency coefficients had the highest precision, with the model predicting R2 being 0.85, the RMSE being 8.11 mg x kg(-1), and RPD being 2.53, indicating the effectiveness of DWT-GA-PLS method in estimating soil alkali hydrolysable nitrogen content.

  20. Genetic transformation of peanut (Arachis hypogaea L.) using cotyledonary node as explant and a promoterless gus::nptII fusion gene based vector

    Indian Academy of Sciences (India)

    T Swathi Anuradha; S K Jami; R S Datla; P B Kirti

    2006-06-01

    We have generated putative promoter tagged transgenic lines in Arachis hypogaea cv JL-24 using cotyledonary node (CN) as an explant and a promoterless gus::nptII bifunctional fusion gene mediated by Agrobacterium transformation. MS medium fortified with 6-benzylaminopurine (BAP) at 4 mg/l in combination with 0.1 mg/l -napthaleneacetic acid (NAA) was the most effective out of the various BAP and NAA combinations tested in multiple shoot bud formation. Parameters enhancing genetic transformation viz. seedling age, Agrobacterium genetic background and co-cultivation periods were studied by using the binary vector p35SGUSINT. Genetic transformation with CN explants from 6-dayold seedlings co-cultivated with Agrobacterium GV2260 strain for 3 days resulted in high kanamycin resistant shoot induction percentage (45%); approximately 31% transformation frequency was achieved with p35S GUSINT in -glucuronidase (GUS) assays. Among the in vivo GUS fusions studied with promoterless gus::nptII construct, GUS-positive sectors occupied 38% of the total transient GUS percentage. We have generated over 141 putative T0 plants by using the promoterless construct and transferred them to the field. Among these, 82 plants survived well in the green house and 5 plants corresponding to 3.54% showed stable integration of the fusion gene as evidenced by GUS, polymerase chain reaction (PCR) and Southern blot analyses. Twenty-four plants were positive for GUS showing either tissue-specific expression or blue spots in at least one plant part. The progeny of 15 T0 plants indicated Mendelian inheritance pattern of segregation for single-copy integration. The tissue-specific GUS expression patterns were more or less similar in both T0 and corresponding T1 progeny plants. We present the differential patterns of GUS expression identified in the putative promoter-tagged transgenic lines in the present communication.

  1. Complex genetic interactions govern the temporal effects of Antennapedia on antenna-to-leg transformations in Drosophila melanogaster

    Indian Academy of Sciences (India)

    Ian Dworkin; Wendy Lee; Fiona Mccloskey; Ellen Larsen

    2007-08-01

    The putative regulatory relationships between Antennapedia (Antp), spalt major (salm) and homothorax (hth) are tested with regard to the sensitive period of antenna-to-leg transformations. Although Antp expression repressed hth as predicted, contrary to expectations, hth did not show increased repression at higher Antp doses, whereas salm, a gene downstream of hth, did show such a dose response. Loss of hth allowed antenna-to-leg transformations but the relative timing of proximal–distal transformations was reversed, relative to transformations induced by ectopic Antp. Finally, overexpression of Hth was only partially able to rescue transformations induced by ectopic Antp. These results indicate that there may be additional molecules involved in antenna/leg identity and that spatial, temporal and dosage relationships are more subtle than suspected and must be part of a robust understanding of molecular network behaviour involved in determining appendage identity in Drosophila melanogaster.

  2. Genetics

    DEFF Research Database (Denmark)

    Christensen, Kaare; McGue, Matt

    2016-01-01

    The sequenced genomes of individuals aged ≥80 years, who were highly educated, self-referred volunteers and with no self-reported chronic diseases were compared to young controls. In these data, healthy ageing is a distinct phenotype from exceptional longevity and genetic factors that protect...

  3. The effect of the number of observations used for Fourier transform infrared model calibration for bovine milk fat composition on the estimated genetic parameters of the predicted data.

    Science.gov (United States)

    Rutten, M J M; Bovenhuis, H; van Arendonk, J A M

    2010-10-01

    Fourier transform infrared spectroscopy is a suitable method to determine bovine milk fat composition. However, the determination of fat composition by gas chromatography, required for calibration of the infrared prediction model, is expensive and labor intensive. It has recently been shown that the number of calibration samples is strongly related to the model's validation r(2) (i.e., accuracy of prediction). However, the effect of the number of calibration samples used, and therefore validation r(2), on the estimated genetic parameters of data predicted using the model needs to be established. To this end, 235 calibration data subsets of different sizes were sampled: n=100, n=250, n=500, and n=1,000 calibration samples. Subsequently, these data subsets were used to calibrate fat composition prediction models for 2 specific fatty acids: C16:0 and C18u (where u=unsaturated). Next, genetic parameters were estimated on predicted fat composition data for these fatty acids. Strong relationships between the number of calibration samples and validation r(2), as well as strong genetic correlations were found. However, the use of n=100 calibration samples resulted in a broad range of validation r(2) values and genetic correlations. Subsequent increases of the number of calibration samples resulted in narrowing patterns for validation r(2) as well as genetic correlations. The use of n=1,000 calibration samples resulted in estimated genetic correlations varying within a range of 0.10 around the average, which seems acceptable. Genetic analyses for the human health-related fatty acids C14:0, C16:0, and C18u, and the ratio of saturated fatty acids to unsaturated fatty acids showed that replacing observations on fat composition determined by gas chromatography by predictions based on infrared spectra reduced the potential genetic gain to 98, 86, 96, and 99% for the 4 fatty acid traits, respectively, in dairy breeding schemes where progeny testing is practiced. We conclude that

  4. Genetic transformation of marine Actinomycete sp. Isolate M048 and expression of a recombinant plasmid carrying the apc gene

    Institute of Scientific and Technical Information of China (English)

    HOU Yanhua; LI Fuchao; QIN Song; WANG Quanfu

    2006-01-01

    Optimal conditions for protoplasts formation of marine Actinomycete sp. isolate M048 were described, dense and disperse mycelia were cultured in SGGP medium, 0.5% glycine, lysozyme exposure (2 mg/cm3, 37 ℃, 40 min), and the concentration of sucrose in protoplast buffer was 0.4 mol/dm3 for keeping the balance of osmotic pressure. Using PEG-mediated protoplasts transformation, the transformation frequency was 89 transformants per microgramme of pIJ702. Meanwhile, an effective transformation procedure was established based on intergeneric conjugation from E. coli ET12567 (pUZ8002) using shuttle vectors pPM801, pPM803 and a(ψ)C31-derived integration vector pIJ8600 containing oriT and attP fragments. Transformation frequencies were 5.30×10-4±0.26×10-4, 8.92×10-4±0.19×10-4 and 6.38×10-5±0.41×10-5, respectively. Further, the heterologous expression of the allophycocyanin gene (apc) in the strain M048 was used to demonstrate this transformation system. SDS-PAGE and Western blot analysis confirmed the expression of recombinant APC (rAPC).

  5. Classification of pumpkin seed oils according to their species and genetic variety by attenuated total reflection Fourier-transform infrared spectroscopy.

    Science.gov (United States)

    Saucedo-Hernández, Yanelis; Lerma-García, María Jesús; Herrero-Martínez, José Manuel; Ramis-Ramos, Guillermo; Jorge-Rodríguez, Elisa; Simí-Alfonso, Ernesto F

    2011-04-27

    Attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR), followed by multivariate treatment of the spectral data, was used to classify seed oils of the genus Cucurbita (pumpkins) according to their species as C. maxima, C. pepo, and C. moschata. Also, C. moschata seed oils were classified according to their genetic variety as RG, Inivit C-88, and Inivit C-2000. Up to 23 wavelength regions were selected on the spectra, each region corresponding to a peak or shoulder. The normalized absorbance peak areas within these regions were used as predictors. Using linear discriminant analysis (LDA), an excellent resolution among all categories concerning both Cucurbita species and C. moschata varieties was achieved. The proposed method was straightforward and quick and can be easily implemented. Quality control of pumpkin seed oils is important because Cucurbita species and genetic variety are both related to the pharmaceutical properties of the oils.

  6. Genetic Transformation of Common Bean (Phaseolus vulgaris L. with the Gus Color Marker, the Bar Herbicide Resistance, and the Barley (Hordeum vulgare HVA1 Drought Tolerance Genes

    Directory of Open Access Journals (Sweden)

    Kingdom Kwapata

    2012-01-01

    Full Text Available Five common bean (Phaseolus vulgaris L. varieties including “Condor,” “Matterhorn,” “Sedona,” “Olathe,” and “Montcalm” were genetically transformed via the Biolistic bombardment of the apical shoot meristem primordium. Transgenes included gus color marker which visually confirmed transgenic events, the bar herbicide resistance selectable marker used for in vitro selection of transgenic cultures and which confirmed Liberty herbicide resistant plants, and the barley (Hordeum vulgare late embryogenesis abundant protein (HVA1 which conferred drought tolerance with a corresponding increase in root length of transgenic plants. Research presented here might assist in production of better P. vulgaris germplasm.

  7. 根癌农杆菌介导灭蚊卵菌贵阳腐霉的遗传转化%The genetic transformation of Pythium guiyangense mediated by Agrobacterium tumefaciens

    Institute of Scientific and Technical Information of China (English)

    赵竟男; 苏晓庆

    2008-01-01

    Pythium guiyangense is a mosquito pathogen,and has been proved to be a promising agent for biological control of mosquitoes.In order to develop the strains adaptable to different ecological environment having stable virulence to mosquito larvae,and being able to prolong the shelf life,an effort was made on transforming the fungus by using homologous or heterologous virulence genes.In this paper,a genetic transformation experiment of P.guiyangense mediated by Agrobacterium tumefaciens is reported.As a result,an A.tumefaciens mediated genetic transformation system was established successfully.

  8. Genetic transformation of Platymonas (Tetraselmis)subcordiformis (Prasinophyceae, Chlorophyta) using particle bombardment and glass-bead agitation

    Institute of Scientific and Technical Information of China (English)

    CUI Yulin; JIANG Peng; WANG Jinfeng; LI Fuchao; CHEN Yingjie; ZHENG Guoting; QIN Song

    2012-01-01

    Platyronas (Tetraselmis) subcordiforris is a unicellular marine green alga.It was found to be very sensitive to the herbicide Basta through a sensitivity test indicating it could be employed as a selective agent.The bar gene is a practicable and selectable marker gene.The vector containing the expression cassette of the bar gene was transferred to P.subcordiformis by both particle bombardment and glass-bead agitation and transformants were then selected using Basta.Finally,Southern blotting analysis indicated that the bar gene had been successfully integrated into the nuclear genome of P.subcordiformis using both of the transgenic techniques,with the transformation efficiency of the glass-bead method being slightly higher than that of particle bombardment.This is the first report on stable transformation ofP.subcordiformis,and will improve fundamental research and enlarge application of this alga.

  9. Evaluation of four Agrobacterium tumefaciens strains for the genetic transformation of tomato (Solanum lycopersicum L.) cultivar Micro-Tom.

    Science.gov (United States)

    Chetty, V J; Ceballos, N; Garcia, D; Narváez-Vásquez, J; Lopez, W; Orozco-Cárdenas, M L

    2013-02-01

    KEY MESSAGE : Agrobacterium tumefaciens strains differ not only in their ability to transform tomato Micro-Tom, but also in the number of transgene copies that the strains integrate in the genome. The transformation efficiency of tomato (Solanum lycopersicum L.) cv. Micro-Tom with Agrobacterium tumefaciens strains AGL1, EHA105, GV3101, and MP90, harboring the plasmid pBI121 was compared. The presence of the nptII and/or uidA transgenes in regenerated T(0) plants was determined by PCR, Southern blotting, and/or GUS histochemical analyses. In addition, a rapid and reliable duplex, qPCR TaqMan assay was standardized to estimate transgene copy number. The highest transformation rate (65 %) was obtained with the Agrobacterium strain GV3101, followed by EHA105 (40 %), AGL1 (35 %), and MP90 (15 %). The mortality rate of cotyledons due to Agrobacterium overgrowth was the lowest with the strain GV3101. The Agrobacterium strain EHA105 was more efficient than GV3101 in the transfer of single T-DNA insertions of nptII and uidA transgenes into the tomato genome. Even though Agrobacterium strain MP90 had the lowest transformation rate of 15 %, the qPCR analysis showed that the strain MP90 was the most efficient in the transfer of single transgene insertions, and none of the transgenic plants produced with this strain had more than two insertion events in their genome. The combination of higher transformation efficiency and fewer transgene insertions in plants transformed using EHA105 makes this Agrobacterium strain optimal for functional genomics and biotechnological applications in tomato.

  10. Organelle transformation.

    Science.gov (United States)

    Bhattacharya, Anjanabha; Kumar, Anish; Desai, Nirali; Parikh, Seema

    2012-01-01

    The source of genetic information in a plant cell is contained in nucleus, plastids, and mitochondria. Organelle transformation is getting a lot of attention nowadays because of its superior performance over the conventional and most commonly used nuclear transformation for obtaining transgenic lines. Absence of gene silencing, strong predictable transgene expression, and its application in molecular pharming, both in pharmaceutical and nutraceuticals, are some of many advantages. Other important benefits of utilizing this technology include the absence of transgene flow, as organelles are maternally inherited. This may increase the acceptability of organelle transformation technology in the development of transgenic crops in a wider scale all over the globe. As the need for crop productivity and therapeutic compounds increases, organelle transformation may be able to bridge the gap, thereby having a definite promise for the future.

  11. 剑麻遗传转化体系的建立与优化%Establishment and Improvement of Agave Genetic Transformation System

    Institute of Scientific and Technical Information of China (English)

    丁静; 徐立; 杜中军; 李志英

    2011-01-01

    Studied the various factors of Agave genetic transformation on the hygromycin(Hyg),bacteria solution concentration,infection time,acetosyringone(As) and carbenicillin(Car).The results showed that the optimal genetic transformation system was as follows: 25 mg/L,infecting in the(OD600=0.6) for 10~15 min.As density was 200 μmol/L.Car density was 400 mg/L.%以剑麻无菌苗叶片为试材,研究了潮霉素(Hyg)、菌液浓度、侵染时间、乙酰丁香酮(As)及羧苄青霉素(Car)等因素对剑麻遗传转化的影响。结果表明,剑麻最适遗传转化体系为:Hyg选择浓度为25 mg/L,OD600=0.6左右的农杆菌菌液,侵染时间为10~15 min,As浓度为200μmol/L,Car浓度为400 mg/L。

  12. Genetic transformation of Slovak cultivar of potato (Solanum tuberosum L.): efficiency and the behavior of the transgene

    NARCIS (Netherlands)

    Moravcikova, J.; Libantova, J.; Matusikova, I.; Libiakova, G.; Nap, J.P.H.; Mlynarova, L.

    2003-01-01

    Transgenic potato plants (Solanum tuberosum L.) of selected Slovak cultivars (cvs.) and one breeding line were obtained by Agrobacterium-mediated transformation. In vitro-grown plants of cvs. Albina, Eta, Malvina, Vila and line 116/86 were tested for their ability to regenerate transgenic plants. In

  13. 共培养环境对玉米遗传转化的影响%Study on Co-culture System to Genetic Transformation of Maize

    Institute of Scientific and Technical Information of China (English)

    王宏伟; 梁业红; 史振声; 张世煌

    2011-01-01

    以携带pCAMBIAl301(含GUS基因)质粒的EHAl05菌株侵染玉米优良自交系齐319幼胚诱导的胚性愈伤组织,共培养5d(特殊试验除外)后,对GUS基因瞬时表达率影响玉米转化的共培养环境进行研究.结果表明,玉米转化效率随共培养基pH降低而升高,临界共培养pH为5.2;共培养基中添加200μmol/L乙酰丁香酮(AS)对提高玉米愈伤组织的转化率是必要的;共培养3和5d的GUS表达率呈现先升高后降低的趋势,以25℃共培养3d的GUS表达率最高,为33.74%.%An optimized transformation system was developed for maize elite inbred lines using Agrobacterium tumefaciens mediated gene transfer by studying various important co-culture systems that affect the efficiency of genetic transformation. The system was performed by transient expression of GUS gene after embryogenic callus co-cultured for five days (except special experiment). The study using the embryogenic callus derived from immature embryos of maize inbred line Qi 319 showed that the genetic transformation efficiency was increased with the decreasing of co-cultivation medium pH. The appropriate pH of co-cultivation medium was 5. 2. It was beneficial to add acetosyringone (AS) into co-cultivation medium for improving transformation efficiency. The result also indicated that the transformation efficiency after adding 200 μmol/L acetosyringone (AS) into the co-cultivation medium was significantly higher than other treatments. Both of the GUS gene expression efficiencies co-cultured for three days and five days were increased at first and then decreased. The highest expression efficiency of GUS gene could reach 33. 74% when co-cultured three days in dark at 25 ℃.

  14. Acetosyringone, pH and temperature effects on transient genetic transformation of immature embryos of Brazilian wheat genotypes by Agrobacterium tumefaciens

    Directory of Open Access Journals (Sweden)

    Ernandes Manfroi

    2015-01-01

    Full Text Available AbstractLow transformation efficiency is one of the main limiting factors in the establishment of genetic transformation of wheat via Agrobacterium tumefaciens. To determine more favorable conditions for T-DNA delivery and explant regeneration after infection, this study investigated combinations of acetosyringone concentration and pH variation in the inoculation and co-cultivation media and co-culture temperatures using immature embryos from two Brazilian genotypes (BR 18 Terena and PF 020037. Based on transient expression of uidA, the most favorable conditions for T-DNA delivery were culture media with pH 5.0 and 5.4 combined with co-culture temperatures of 22 °C and 25 °C, and a 400 μM acetosyringone supplement. These conditions resulted in blue foci in 81% of the embryos. Media with more acidic pH also presented reduced A. tumefaciensovergrowth during co-culture, and improved regeneration frequency of the inoculated explants. BR 18 Terena was more susceptible to infection by A. tumefaciens than PF 020037. We found that it is possible to improve T-DNA delivery and explant regeneration by adjusting factors involved in the early stages of A. tumefaciens infection. This can contribute to establishing a stable transformation procedure in the future.

  15. [Construction of a bivalent plant expression vector carrying VvSUC11 and VvSUC12 genes and its genetic transformation in sugar beet].

    Science.gov (United States)

    Yin, Donglin; Zhu, Jianbo; Wang, Aiying; Xiang, Benchun

    2011-08-01

    We have recombined genes VvSUC11, VvSUC12 from Vitis vinifera L., and root-specific promoters of sweet potato storage protein gene from Ipomoea batatas L. Lam., named as SP1 and SP2. We have constructed a vector pCAMBIA2301-SP1- VvSUC11-SP2-VvSUC12 using pCAMBIA2301 as an original vector. VvSUC11 and VvSUC12 were under the control of root-specific promoters of sweet potato storage protein gene. We transformed the vector into KWS-9103 breeding line of Beta vulgaris L. with Agrobacterium-mediated transformation. We have established the optimal genetic transformation protocol of sugar beet as following: the explants pre-cultured for 4 days were immersed in Agrobacterium suspension of OD(600)=0.5, supplemented with 0.005% Silwet L-77, and followed by a 4-day culture on medium containing cefotaxime, then the buds were selected on medium containing kanamycin and cefotaxime. The percentage of kanamycin-resistant buds was as high as 42%. Results of PCR and RT-PCR proved that the target genes had integrated into sugar beet genome and expressed. It will lay a foundation for further studying their function in Beta vulgaris.

  16. Study on the Maize Genetic Transformation Technology%转基因玉米的研究进展

    Institute of Scientific and Technical Information of China (English)

    于海波; 张玲

    2011-01-01

    Since 1996, genetically modified crops had been large-scale commercial cultivation and developed rapidly in the international debate environment. Nowadays, it has been the fastest growing crop of new technology in modern history and created a huge economic, social and environmental benefit. The research and application of genetically modified maize has made maize breeding to a new era, and has opened up broad prospects for the cultivation of new varieties of maize. The domestic and international commercialization of genetically modified maize overview and trends were reviewed in this paper, and pointed out the direction for future of genetically modified maize industry.%转基因农作物自1996年大规模商业化种植以来,在国际争论的环境中迅猛发展,成为近代历史上应用最快的作物新技术,创造了巨大的经济、社会与环境效益.转基因玉米研究与应用使玉米的育种工作进入到新时代,为培育玉米新品种开辟了广阔的前景.综述国内外转基因玉米的商业化概况和发展趋势,为今后转基因玉米产业化方向提供参考.

  17. Enhancement of lipid productivity in oleaginous Colletotrichum fungus through genetic transformation using the yeast CtDGAT2b gene under model-optimized growth condition.

    Directory of Open Access Journals (Sweden)

    Prabuddha Dey

    Full Text Available Oleaginous fungi are of special interest among microorganisms for the production of lipid feedstocks as they can be cultured on a variety of substrates, particularly waste lingocellulosic materials, and few fungal strains are reported to accumulate inherently higher neutral lipid than bacteria or microalgae. Previously, we have characterized an endophytic filamentous fungus Colletotrichum sp. DM06 that can produce total lipid ranging from 34% to 49% of its dry cell weight (DCW upon growing with various carbon sources and nutrient-stress conditions. In the present study, we report on the genetic transformation of this fungal strain with the CtDGAT2b gene, which encodes for a catalytically efficient isozyme of type-2 diacylglycerol acyltransferase (DGAT from oleaginous yeast Candida troplicalis SY005. Besides the increase in size of lipid bodies, total lipid titer by the transformed Colletotrichum (lipid content ∼73% DCW was found to be ∼1.7-fold more than the wild type (lipid content ∼38% DCW due to functional activity of the CtDGAT2b transgene when grown under standard condition of growth without imposition of any nutrient-stress. Analysis of lipid fractionation revealed that the neutral lipid titer in transformants increased up to 1.8-, 1.6- and 1.5-fold compared to the wild type when grown under standard, nitrogen stress and phosphorus stress conditions, respectively. Lipid titer of transformed cells was further increased to 1.7-fold following model-based optimization of culture conditions. Taken together, ∼2.9-fold higher lipid titer was achieved in Colletotrichum fungus due to overexpression of a rate-limiting crucial enzyme of lipid biosynthesis coupled with prediction-based bioprocess optimization.

  18. Genetic transformation of Indian bread (T. aestivum and pasta (T. durum wheat by particle bombardment of mature embryo-derived calli

    Directory of Open Access Journals (Sweden)

    Khurana Paramjit

    2003-09-01

    Full Text Available Abstract Background Particle bombardment has been successfully employed for obtaining transgenics in cereals in general and wheat in particular. Most of these procedures employ immature embryos which are not available throughout the year. The present investigation utilizes mature seeds as the starting material and the calli raised from the hexaploid Triticum aestivum and tetraploid Triticum durum display a high regeneration response and were therefore used as the target tissue for genetic transformation by the biolistic approach. Results Mature embryo-derived calli of bread wheat (Triticum aestivum, cv. CPAN1676 and durum wheat (T. durum, cv. PDW215 were double bombarded with 1.1 gold microprojectiles coated with pDM302 and pAct1-F at a target distance of 6 cm. Southern analysis using the bar gene as a probe revealed the integration of transgenes in the T0 transformants. The bar gene was active in both T0 and T1 generations as evidenced by phosphinothricin leaf paint assay. Approximately 30% and 33% primary transformants of T. aestivum and T. durum, respectively, were fertile. The transmission of bar gene to T1 progeny was demonstrated by PCR analysis of germinated seedlings with primers specific to the bar gene. Conclusions The transformation frequency obtained was 8.56% with T. aestivum and 10% with T. durum. The optimized protocol was subsequently used for the introduction of the barley gene encoding a late embryogenesis abundant protein (HVA1 in T. aestivum and T. durum. The presence of the HVA1 transgene was confirmed by Southern analysis in the T0 generation in case of Triticum aestivum, and T0 and T1 generation in Triticum durum.

  19. Enhancement of lipid productivity in oleaginous Colletotrichum fungus through genetic transformation using the yeast CtDGAT2b gene under model-optimized growth condition.

    Science.gov (United States)

    Dey, Prabuddha; Mall, Nikunj; Chattopadhyay, Atrayee; Chakraborty, Monami; Maiti, Mrinal K

    2014-01-01

    Oleaginous fungi are of special interest among microorganisms for the production of lipid feedstocks as they can be cultured on a variety of substrates, particularly waste lingocellulosic materials, and few fungal strains are reported to accumulate inherently higher neutral lipid than bacteria or microalgae. Previously, we have characterized an endophytic filamentous fungus Colletotrichum sp. DM06 that can produce total lipid ranging from 34% to 49% of its dry cell weight (DCW) upon growing with various carbon sources and nutrient-stress conditions. In the present study, we report on the genetic transformation of this fungal strain with the CtDGAT2b gene, which encodes for a catalytically efficient isozyme of type-2 diacylglycerol acyltransferase (DGAT) from oleaginous yeast Candida troplicalis SY005. Besides the increase in size of lipid bodies, total lipid titer by the transformed Colletotrichum (lipid content ∼73% DCW) was found to be ∼1.7-fold more than the wild type (lipid content ∼38% DCW) due to functional activity of the CtDGAT2b transgene when grown under standard condition of growth without imposition of any nutrient-stress. Analysis of lipid fractionation revealed that the neutral lipid titer in transformants increased up to 1.8-, 1.6- and 1.5-fold compared to the wild type when grown under standard, nitrogen stress and phosphorus stress conditions, respectively. Lipid titer of transformed cells was further increased to 1.7-fold following model-based optimization of culture conditions. Taken together, ∼2.9-fold higher lipid titer was achieved in Colletotrichum fungus due to overexpression of a rate-limiting crucial enzyme of lipid biosynthesis coupled with prediction-based bioprocess optimization.

  20. Acute leukemia viruses E26 and avian myeloblastosis virus have related transformation-specific RNA sequences but different genetic structures, gene products, and oncogenic properties

    Science.gov (United States)

    Bister, Klaus; Nunn, Michael; Moscovici, Carlo; Perbal, Bernard; Baluda, Marcel A.; Duesberg, Peter H.

    1982-01-01

    Replication-defective acute leukemia viruses E26 and myeloblastosis virus (AMV) cause distinct leukemias although they belong to the same subgroup of oncogenic avian tumor viruses based on shared transformation-specific (onc) RNA sequences. E26 causes predominantly erythroblastosis in chicken and in quail, whereas AMV induces a myeloid leukemia. However, upon cultivation in vitro for >1 month, a majority of surviving hemopoietic cells of E26-infected animals bear myeloid markers similar to those of AMV-transformed cells. We have analyzed the genetic structure and gene products of E26 virus for a comparison with those of AMV. An E26/helper virus complex was found to contain two RNA species: a 5.7-kilobase (kb) RNA that hybridizes with cloned AMV-specific proviral DNA and hence is probably the E26 genome; and an 8.5-kb RNA that is unrelated to AMV and represents helper virus RNA. Thus, E26 RNA is smaller than 7.5-kb AMV RNA. Hybridization of size-selected poly(A)-terminating E26 RNA fragments with AMV-specific DNA indicated that the shared specific sequences are located in the 5′ half of the E26 genome as opposed to a 3′ location in AMV RNA. In nonproducer cells transformed in vitro by E26, a gag-related nonstructural 135,000-dalton protein (p135) was found. No gag(Pr76) or gag-pol (Pr180) precursors of essential virion proteins, which are present in AMV nonproducer cells, were observed. p135 was also found in cultured E26 virus producing cells of several leukemic chickens, and its intracellular concentration relative to that of the essential virion proteins encoded by the helper virus correlates with the ratio of E26 to helper RNA in virions released by these cells. p135 is phosphorylated but not glycosylated; antigenically it is not related to the pol or env gene products. It appears to be coded for by a partial gag gene and by E26-specific RNA sequences, presumably including those shared with AMV. Hence, AMV and E26 appear to use different strategies for the

  1. Genetic components of milk Fourier-transform infrared spectra used to predict breeding values for milk composition and quality traits in dairy goats.

    Science.gov (United States)

    Dagnachew, B S; Meuwissen, T H E; Adnøy, T

    2013-09-01

    The usual practice today is that milk component phenotypes are predicted using Fourier-transform infrared (FTIR) spectra and they are then, together with pedigree information, used in BLUP for calculation of individual estimated breeding values. Here, this is referred to as the indirect prediction (IP) approach. An alternative approach-a direct prediction (DP) method-is proposed, where genetic analyses are directly conducted on the milk FTIR spectral variables. Breeding values of all derived milk traits (protein, fat, fatty acid composition, and coagulation properties, among others) can then be predicted as traits correlated only to the genetic information of the spectra. For the DP, no need exists to predict the phenotypes before calculating breeding values for each of the traits-the genetic analysis is done once for the spectra, and is applicable to all traits derived from the spectra. The aim of the study was to compare the effects of DP and IP of milk composition and quality traits on prediction error variance (PEV) and genetic gain. A data set containing 27,927 milk FTIR spectral observations and milk composition phenotypes (fat, lactose, and protein) belonging to 14,869 goats of 271 herds was used for training and evaluating models. Partial least squares regression was used for calibrating prediction models for fat, protein, and lactose percentages. Restricted maximum likelihood was used to estimate variance components of the spectral variables after principal components analysis was applied to reduce the spectral dimension. Estimated breeding values were predicted for fat, lactose, and protein percentages using DP and IP methods. The DP approach reduced the mean PEV by 3.73, 4.07, and 7.04% for fat, lactose, and protein percentages, respectively, compared with the IP method. Given the reduction in PEV, relative genetic gains were 2.99, 2.78, and 4.85% for fat, lactose, and protein percentages, respectively. We concluded that more accurate estimated breeding

  2. Contrasted Fitness Costs of Docking and Antibacterial Constructs in the EE and EVida3 Strains Validates Two-Phase Anopheles gambiae Genetic Transformation System.

    Directory of Open Access Journals (Sweden)

    Doug Paton

    Full Text Available The deployment of transgenic mosquitoes carrying genes for refractoriness to malaria has long been seen as a futuristic scenario riddled with technical difficulties. The integration of anti-malarial effector genes and a gene-drive system into the mosquito genome without affecting mosquito fitness is recognized as critical to the success of this malaria control strategy. Here we conducted detailed fitness studies of two Anopheles gambiae s.s. transgenic lines recently developed using a two-phase targeted genetic transformation system. In replicated cage-invasion experiments, males and females of the EE Phase-1 docking strain and EVida3 Phase-2 strain loaded with an antimicrobial peptide (AMP expressed upon blood-feeding, were mixed with individuals of a recently-colonized strain of the Mopti chromosomal form. The experimental design enabled us to detect initial strain reproductive success differences, assortative mating and hybrid vigor that may characterize mosquito release situations. In addition, the potential fitness costs of the unloaded Phase-1 and loaded Phase-2 genetic constructs, independent of the strains' original genetic backgrounds, were estimated between the 1(st instar larvae, pupae and adult stages over 10 generations. The Phase-1 unloaded docking cassette was found to have significantly lower allelic fitness relative to the wild type allele during larval development. However, overall genotypic fitness was comparable to the wild type allele across all stages leading to stable equilibrium in all replicates. In contrast, the Phase-2 construct expressing EVida3 disappeared from all replicates within 10 generations due to lower fitness of hemi- and homozygous larvae, suggesting costly background AMP expression and/or of the DsRed2 marker. This is the first study to effectively partition independent fitness stage-specific determinants in unloaded and loaded transgenic strains of a Phase-1-2 transformation system. Critically, the high

  3. Contrasted Fitness Costs of Docking and Antibacterial Constructs in the EE and EVida3 Strains Validates Two-Phase Anopheles gambiae Genetic Transformation System.

    Science.gov (United States)

    Paton, Doug; Underhill, Anne; Meredith, Janet; Eggleston, Paul; Tripet, Frederic

    2013-01-01

    The deployment of transgenic mosquitoes carrying genes for refractoriness to malaria has long been seen as a futuristic scenario riddled with technical difficulties. The integration of anti-malarial effector genes and a gene-drive system into the mosquito genome without affecting mosquito fitness is recognized as critical to the success of this malaria control strategy. Here we conducted detailed fitness studies of two Anopheles gambiae s.s. transgenic lines recently developed using a two-phase targeted genetic transformation system. In replicated cage-invasion experiments, males and females of the EE Phase-1 docking strain and EVida3 Phase-2 strain loaded with an antimicrobial peptide (AMP) expressed upon blood-feeding, were mixed with individuals of a recently-colonized strain of the Mopti chromosomal form. The experimental design enabled us to detect initial strain reproductive success differences, assortative mating and hybrid vigor that may characterize mosquito release situations. In addition, the potential fitness costs of the unloaded Phase-1 and loaded Phase-2 genetic constructs, independent of the strains' original genetic backgrounds, were estimated between the 1(st) instar larvae, pupae and adult stages over 10 generations. The Phase-1 unloaded docking cassette was found to have significantly lower allelic fitness relative to the wild type allele during larval development. However, overall genotypic fitness was comparable to the wild type allele across all stages leading to stable equilibrium in all replicates. In contrast, the Phase-2 construct expressing EVida3 disappeared from all replicates within 10 generations due to lower fitness of hemi- and homozygous larvae, suggesting costly background AMP expression and/or of the DsRed2 marker. This is the first study to effectively partition independent fitness stage-specific determinants in unloaded and loaded transgenic strains of a Phase-1-2 transformation system. Critically, the high fitness of the

  4. Progress of Methodology of Genetic Transformation of Banana%香蕉遗传转化方法研究进展

    Institute of Scientific and Technical Information of China (English)

    黄永红; 易干军; 周碧容; 曾继吾; 吴元立

    2006-01-01

    Banana is an important staple food and source of carbohydrates in developing counties of tropical and subtropical. But many pests and diseases have significantly affected banana cultivation. Most cultivars of banana are triploid, long life cycle, and sterility, which resulted in difficulty for its reproduction and breeding. In order to conquer limitations of conventional breeding and to improve cultivars qualities, transgenic approaches have been introduced. The development of transgenic Musa plants has been achieved using electroporation, microprojectile bombardment procedure, and A grobacterium-mediated transformation etc. The successful application of A grobacterium- mediated transformation protocol is a great breakthrough in the development of banana cultivars improvement. The explants of the genetic transformation of Musa have developed into protoplast, embryogenic cell suspensions, meristematic tissues and thin cell layer. Presently, with the advancement of biology, some more efficient genetic transformation methods including integrated bombardment and Agrobacterium transformation, centrifugation-assisted Agrobacterium-mediated transformation (CAAT) protocol, and vacuum infiltration technique have been developed and made great progress, which will accelerate the progress of banana breeding.%香蕉是热带亚热带发展中国家重要的粮食作物和碳水化合物来源.但近年来,香蕉生产受到严重的病虫危害.大多数香蕉栽培品种是三倍体,生长周期长,而且不孕.由于没有种子,给繁殖和育种带来一定的困难.遗传转化技术的发展为香蕉品种的改良提供了一种有效的手段.香蕉的遗传转化方法有电激法、基因枪法、农杆菌介导法等.农杆菌介导法的应用是香蕉品种改良的一个重大突破.香蕉遗传转化的外植体也发展到多种,有原生质体,胚性细胞悬浮系,分生组织,以及横切薄片等.近几年,随着分子生物学的发展,出现了转化效

  5. Adventitious shoot regeneration of Platanus acerifolia Willd.facilitated by Timentin, an antibiotic for suppression of Agrobacterium tumefaciens in genetic transformation

    Institute of Scientific and Technical Information of China (English)

    Li Zhi-neng; Liu Guo-feng; Fang Fang; Bao Man-zhu

    2007-01-01

    The effects of Timentin and cefotaxime (Cef) on shoot regeneration of the London plane tree (Platanus acerifolia Willd.) and their use for the suppression of Agrobacterium tumefaciens in Agrobacterium-mediated genetic transformation were compared.Shoot regeneration was significantly reduced on the media with Cef at concentrations from 100 to 500 mg·L-1. Timentin showed negative effect on plant regeneration at concentrations of 100 and 500 mg·L-1; however, 300 mg·L-1 Timentin was shown to facilitate shoot regeneration significantly and the regeneration frequency increased from 64% (control) to 88%. Effective suppression of A.tumefaciens could be obtained with 500 mg·L-1 Cef, but plant regeneration was completely inhibited at this level. The A. tumefaciens on infected P. acerifolia leaf tissues was visually undetectable after three subcultures on a medium with 300 mg·L-1 Timentin. Considering the effect of Cef and Timentin on plant regeneration and suppression of Agrobacteria, Timentin at 300 mg·L-1 is the preferred application in A. tumefaciens-mediated transformation of P.acerifolia.

  6. [Genetic transformation of buckwheat ( Fagopyrum esculentum Moench ) with AtNHX1 gene and regeneration of salt-tolerant transgenic plants].

    Science.gov (United States)

    Cheng, Li-Hong; Zhang, Bo; Xu, Zi-Qin

    2007-01-01

    The Arabidopsis thaliana tonoplast Na+ /H+ antiporter gene, AtNHX1, was transferred into buckwheat by Agrobacterium-mediated method. Transgenic buckwheat plants were regenerated and selected on MS basal medium supplemented with 2.0mg/L 6-BA, 1.0mg/L KT, 0.lmg/L IAA, 50mg/L kanamycin and 500mg/L carbenicillin. 426 seedlings from 36 resistant calli originated from 864 explants (transformed about at 4.17 percentage) exhibited resistance to kanamycin. The transformants were confirmed by PCR, Southern blotting, RT-PCR and Northern blotting analysis. After stress treatment for 6 weeks with 200mmol/L NaCl, transgenic plants survived, while wild-type plants did not. After 3 days of stress treatment through different concentrations of NaCl, transgenic plants accumulated higher concentration of Na+ and proline than the control plants. However, the K+ concentration of transgenic plants declined in comparison with the control plants. Moreover, the rutin content of the roots, stems and leaves of transgenic buckwheat increased than those of the control plants. These results showed that it could be possible to improve the salt-tolerance of crops with genetic technology.

  7. Genetic transformation of an obligate anaerobe, P. gingivalis for FMN-green fluorescent protein expression in studying host-microbe interaction.

    Directory of Open Access Journals (Sweden)

    Chul Hee Choi

    Full Text Available The recent introduction of "oxygen-independent" flavin mononucleotide (FMN-based fluorescent proteins (FbFPs is of major interest to both eukaryotic and prokaryotic microbial biologists. Accordingly, we demonstrate for the first time that an obligate anaerobe, the successful opportunistic pathogen of the oral cavity, Porphyromonas gingivalis, can be genetically engineered for expression of the non-toxic green FbFP. The resulting transformants are functional for studying dynamic bacterial processes in living host cells. The visualization of the transformed P. gingivalis (PgFbFP revealed strong fluorescence that reached a maximum emission at 495 nm as determined by fluorescence microscopy and spectrofluorometry. Human primary gingival epithelial cells (GECs were infected with PgFbFP and the bacterial invasion of host cells was analyzed by a quantitative fluorescence microscopy and antibiotic protection assays. The results showed similar levels of intracellular bacteria for both wild type and PgFbFP strains. In conjunction with organelle specific fluorescent dyes, utilization of the transformed strain provided direct and accurate determination of the live/metabolically active P. gingivalis' trafficking in the GECs over time. Furthermore, the GECs were co-infected with PgFbFP and the ATP-dependent Clp serine protease-deficient mutant (ClpP- to study the differential fates of the two strains within the same host cells. Quantitative co-localization analyses displayed the intracellular PgFbFP significantly associated with the endoplasmic reticulum network, whereas the majority of ClpP- organisms trafficked into the lysosomes. Hence, we have developed a novel and reliable method to characterize live host cell-microbe interactions and demonstrated the adaptability of FMN-green fluorescent protein for studying persistent host infections induced by obligate anaerobic organisms.

  8. The role of IREB2 and transforming growth factor beta-1 genetic variants in COPD: a replication case-control study

    LENUS (Irish Health Repository)

    Chappell, Sally L

    2011-02-14

    Abstract Background Genetic factors are known to contribute to COPD susceptibility and these factors are not fully understood. Conflicting results have been reported for many genetic studies of candidate genes based on their role in the disease. Genome-wide association studies in combination with expression profiling have identified a number of new candidates including IREB2. A meta-analysis has implicated transforming growth factor beta-1 (TGFbeta1) as a contributor to disease susceptibility. Methods We have examined previously reported associations in both genes in a collection of 1017 white COPD patients and 912 non-diseased smoking controls. Genotype information was obtained for seven SNPs in the IREB2 gene, and for four SNPs in the TGFbeta1 gene. Allele and genotype frequencies were compared between COPD cases and controls, and odds ratios were calculated. The analysis was adjusted for age, sex, smoking and centre, including interactions of age, sex and smoking with centre. Results Our data replicate the association of IREB2 SNPs in association with COPD for SNP rs2568494, rs2656069 and rs12593229 with respective adjusted p-values of 0.0018, 0.0039 and 0.0053. No significant associations were identified for TGFbeta1. Conclusions These studies have therefore confirmed that the IREB2 locus is a contributor to COPD susceptibility and suggests a new pathway in COPD pathogenesis invoking iron homeostasis.

  9. Method for Optimal Sensor Deployment on 3D Terrains Utilizing a Steady State Genetic Algorithm with a Guided Walk Mutation Operator Based on the Wavelet Transform

    Science.gov (United States)

    Unaldi, Numan; Temel, Samil; Asari, Vijayan K.

    2012-01-01

    One of the most critical issues of Wireless Sensor Networks (WSNs) is the deployment of a limited number of sensors in order to achieve maximum coverage on a terrain. The optimal sensor deployment which enables one to minimize the consumed energy, communication time and manpower for the maintenance of the network has attracted interest with the increased number of studies conducted on the subject in the last decade. Most of the studies in the literature today are proposed for two dimensional (2D) surfaces; however, real world sensor deployments often arise on three dimensional (3D) environments. In this paper, a guided wavelet transform (WT) based deployment strategy (WTDS) for 3D terrains, in which the sensor movements are carried out within the mutation phase of the genetic algorithms (GAs) is proposed. The proposed algorithm aims to maximize the Quality of Coverage (QoC) of a WSN via deploying a limited number of sensors on a 3D surface by utilizing a probabilistic sensing model and the Bresenham's line of sight (LOS) algorithm. In addition, the method followed in this paper is novel to the literature and the performance of the proposed algorithm is compared with the Delaunay Triangulation (DT) method as well as a standard genetic algorithm based method and the results reveal that the proposed method is a more powerful and more successful method for sensor deployment on 3D terrains. PMID:22666078

  10. Congenic mice provide in vivo evidence for a genetic locus that modulates intrinsic transforming growth factor β1-mediated signaling and bone acquisition.

    Science.gov (United States)

    Mukherjee, Aditi; Larson, Emily A; Carlos, Amy S; Belknap, John K; Rotwein, Peter; Klein, Robert F

    2012-06-01

    Osteoporosis, the most common skeletal disorder, is characterized by low bone mineral density (BMD) and an increased risk of fragility fractures. BMD is the best clinical predictor of future osteoporotic fracture risk, but is a complex trait controlled by multiple environmental and genetic determinants with individually modest effects. Quantitative trait locus (QTL) mapping is a powerful method for identifying chromosomal regions encompassing genes involved in shaping complex phenotypes, such as BMD. Here we have applied QTL analysis to male and female genetically-heterogeneous F(2) mice derived from a cross between C57BL/6 and DBA/2 strains, and have identified 11 loci contributing to femoral BMD. Further analysis of a QTL on mouse chromosome 7 following the generation of reciprocal congenic strains has allowed us to determine that the high BMD trait, which tracks with the DBA/2 chromosome and exerts equivalent effects on male and female mice, is manifested by enhanced osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro and by increased growth of metatarsal bones in short-term primary culture. An insertion/deletion DNA polymorphism in Ltbp4 exon 12 that causes the in-frame removal of 12 codons in the DBA/2-derived gene maps within 0.6 Mb of the marker most tightly linked to the QTL. LTBP4, one of four paralogous mouse proteins that modify the bioavailability of the transforming growth factor β (TGF-β) family of growth factors, is expressed in differentiating MSC-derived osteoblasts and in long bones, and reduced responsiveness to TGF-β1 is observed in MSCs of mice homozygous for the DBA/2 chromosome 7. Taken together, our results identify a potential genetic and biochemical relationship between decreased TGF-β1-mediated signaling and enhanced femoral BMD that may be regulated by a variant LTBP4 molecule. Copyright © 2012 American Society for Bone and Mineral Research.

  11. Plant Regeneration from Immature Inflorescence Culture and Genetic Transformation of Wheatgrass (Agropyron cristatum × A. Desertorum cv. Hycrest-Mengnong)

    Institute of Scientific and Technical Information of China (English)

    HUO Xiu-wen; WEI Jian-hua; XU Chun-bo; MI Fu-gui; YUN Jin-feng

    2006-01-01

    The plants of hybrid wheatgrass (A. Cristatum ×A. Desertorum cv. Hycrest-Mengnong) were directly induced from embryogenic callus regenerated from immature inflorescence. Immature inflorescence was cultured on improved MS medium containing 2.0-3.0 mg L-1 2,4-D to regenerate callus. The calli were then transferred to hormone-free MS medium for differentiation and 1/2 MS medium for rooting. Results showed that callus initiation frequency was 83.4% and plant regeneration frequency was 59.6%. Phosphinothricin acetyltransferase (bar) gene was transformed into the hybrid wheatgrass by particle bombardment. Resistant callus was obtained using selecting agent, herbicide glufosinate of 0.5 mg L-1, and some transgenic plants were recovered in vitro. The transgenic plants were identified by PCR and Southern blot analysis and these plants developed normally in the glufosinate medium, whereas the nontransgenic plants did not. The results demonstrated that bar cDNA integrated into the genomic DNA of the transgenic plants. The transgenic frequencies of bar gene were 1.1%.

  12. Transforming growth factor-β genetic polymorphisms on development of liver cirrhosis in a meta-analysis.

    Science.gov (United States)

    Wu, Xiao-Dan; Zeng, Kai; Gong, Can-Sheng; Chen, Jinhua; Chen, Yan-Qing

    2013-01-01

    Transforming growth factor-β (TGF-β) protein has been supposed to be a risk factor for liver cirrhosis; however, the associations between its genes (TGF-β -509C>T and +869T>C) and liver cirrhosis remained unclear. This study was to quantitatively analyze the correlations by using a meta-analysis. Pubmed, Embase, Wanfang databases were retrieved up to November 1st, 2011. Odds ratio (OR) and 95 % confidence interval (95 %CI) were used to demonstrate the strength of association, and P T polymorphism, and eight studies were for +869T>C polymorphism. Combined results indicated that neither TGF-β -509C>T nor +869T>C polymorphisms were associated with risk of liver cirrhosis [OR (95 % CI): 0.79 (0.60-1.04) for CT vs. TT of -509C>T and 0.87 (0.68-1.12) for CT vs. CC of +869T>C], with no between-study heterogeneity. In addition, subgroups analyses still inferred that two polymorphisms were not associated with risk of liver cirrhosis for HBV-infected patients, Asians and for Population-based studies. This meta-analysis indicated that neither TGF-β -509C>T nor +869T>C polymorphisms were associated with risk of liver cirrhosis, regardless of HBV infection or not.

  13. A Two-dimensional Genetic Algorithm Based on the Eno-Haar Wavelet Transform%一种基于Eno-Haar小波变换二维遗传算法

    Institute of Scientific and Technical Information of China (English)

    宋锦萍; 赵晨萍; 李登峰

    2007-01-01

    A two-dimensional genetic algorithm of wavelet coefficient is presented by using the ENO wavelet transform and the decomposed characterization of the two-dimensional Haar wavelet. And simulated by the ENO interpolation the article shows the affectivity and the superiority of this algorithm.

  14. Genetic analysis of the Fourier-transform infrared spectra of bovine milk with emphasis on individual wavelengths related to specific chemical bonds.

    Science.gov (United States)

    Bittante, G; Cecchinato, A

    2013-09-01

    Fourier-transform infrared (FTIR) spectra are used to predict the fat, protein, casein, and lactose contents of milk. These estimates are currently used to predict the individual estimated breeding values of animals. The objective of the present study was to estimate the genetic variation and heritabilities of the milk transmittance spectrum at each individual FTIR wave. Milk was sampled once per cow from a total of 1,064 Italian Brown Swiss cows from 30 herds, sired by 50 artificial insemination sires. The FTIR spectra of all samples were collected within 3 h of sampling from 25 mL of milk. The obtained spectral range comprised wavenumbers 5,000 to 930×cm(-1), corresponding to wavelengths 2.00 to 10.76 μm and frequencies from 149.9 to 27.9 THz, for a total of 1,056 waves. These were acquired using a MilkoScan FT120 FTIR interferometer (Foss Electric A/S, Hillerød, Denmark). Each spectral data point was treated as a single trait and analyzed using an animal model REML method. The results indicated that the transmittance of the bovine milk FTIR spectrum was heritable for most individual waves in the wavenumber interval from 5,000 to 930×cm(-1). Moreover, the transmittance of contiguous FTIR waves was much more highly correlated in terms of the average value and phenotypic variation, compared with genetic variation. In the present study, we characterized 5 regions of the FTIR spectrum that were relevant to the analysis of milk; 2 regions, one in the transition area between the short-wavelength infrared (SWIR) and mid-wavelength infrared (MWIR) divisions of the electromagnetic spectrum (SWIR-MWIR region) and another very short region in the MWIR division (MWIR-2 region), were characterized by very high phenotypic variability in the transmittance of individual milk samples within each wave. This was caused by the absorption peaks of water, which can mask the effects of other important milk components. These regions also showed high genetic variability in

  15. Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines

    Directory of Open Access Journals (Sweden)

    Neumann Elisabeth

    2006-01-01

    Full Text Available Abstract Background The use of lactic acid bacteria as vehicles to delivery antigens to immunize animals is a promising issue. When genetically modified, these bacteria can induce a specific local and systemic immune response against selected pathogens. Gastric acid and bile salts tolerance, production of antagonistic substances against pathogenic microorganisms, and adhesive ability to gut epithelium are other important characteristics that make these bacteria useful for oral immunization. Results Bacteria isolated on de Man, Rogosa and Sharpe medium (MRS from different gastrointestinal portions of broiler chicks were evaluated for their resistance to artificial gastric acid and bile salts, production of hydrogen peroxide, and cell surface hydrophobicity. Thirty-eight isolates were first typed at species level by PCR amplification of 16S-23S rRNA intergenic spacers using universal primers that anneal within 16S and 23S genes, followed by restriction digestion analyses of PCR amplicons (PCR-ARDRA. An expression cassette was assembled onto the pCR2.1-Topo vector by cloning the promoter, leader peptide, cell wall anchor and terminator sequences derived from the laminin binding S-layer protein gene of L. crispatus strain F5.7 (lbs gene. A sequence encoding the green fluorescent protein (GFP was inserted as reporter gene, and an erythromycin resistance gene was added as selective marker. All constructs were able to express GFP in the cloning host E. coli XL1-Blue and different Lactobacillus strains as verified by FACS and laser scanning confocal microscopy. Conclusion Lactobacillus isolated from gastrointestinal tract of broiler chickens and selected for probiotic characteristics can be genetically modified by introducing an expression cassette into the lbs locus. The transformed bacteria expressed on its cell wall surface different fluorescent proteins used as reporters of promoter function. It is possible then that similar bacterial model

  16. 柚木cry1A(b)基因的遗传转化%Genetic transformation of cry1A(b) gene into teak

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Teak (Tectona grandis) provides one of the most highly sought after timber in the world and is a widely recommended species for reforestation. As such teak is widely planted in Malaysia. Though no serious outbreaks have been recorded for teak in Malaysia, but insect attack remains the most important threat to the timber industry. Thus, in efforts to overcome the problem, an integrated pest management system needs to be developed. Spraying of commercial Bt has been a common practice in addressing minor outbreaks. However, one of the main limitations of the spraying technique is poor coverage, especially on plant surfaces. Poor coverage, however, could be overcome by planting insect resistant trees. In addition, the approach of using genetic engineering in addressing the above problem proves to be possible with the advancement made in genetic transformation of trees especially in the last decade. This, together with improved knowledge on gene function following improved DNA recombinant techniques promises the major advancement in pest management of forest species. This report demonstrates the possibility of transferring foreign gene into teak cells. In this study, nodal segments of teak were subjected to particle bombardment. Nodal segments bombarded with gold particles coated with plasmid DNA carrying hygromycin phosphotransferase (hpt), β-glucuronidase (gus) and cry1A(b) genes were then transferred onto medium for shoot development. The shoots were than transferred onto the same medium supplemented with 10mg/L hygromycin for selection. Selection was repeated several times with six-week subculture intervals on the same Hm-containing media. The presence of the transgenes in the Hmr plants was confirmed using PCR.

  17. Genetic transformation of novel isolates of chicken Lactobacillus bearing probiotic features for expression of heterologous proteins: a tool to develop live oral vaccines

    Science.gov (United States)

    Mota, Rodrigo M; Moreira, João Luiz S; Souza, Marcelo R; Fátima Horta, M; Teixeira, Santuza MR; Neumann, Elisabeth; Nicoli, Jacques R; Nunes, Álvaro C

    2006-01-01

    Background The use of lactic acid bacteria as vehicles to delivery antigens to immunize animals is a promising issue. When genetically modified, these bacteria can induce a specific local and systemic immune response against selected pathogens. Gastric acid and bile salts tolerance, production of antagonistic substances against pathogenic microorganisms, and adhesive ability to gut epithelium are other important characteristics that make these bacteria useful for oral immunization. Results Bacteria isolated on de Man, Rogosa and Sharpe medium (MRS) from different gastrointestinal portions of broiler chicks were evaluated for their resistance to artificial gastric acid and bile salts, production of hydrogen peroxide, and cell surface hydrophobicity. Thirty-eight isolates were first typed at species level by PCR amplification of 16S-23S rRNA intergenic spacers using universal primers that anneal within 16S and 23S genes, followed by restriction digestion analyses of PCR amplicons (PCR-ARDRA). An expression cassette was assembled onto the pCR2.1-Topo vector by cloning the promoter, leader peptide, cell wall anchor and terminator sequences derived from the laminin binding S-layer protein gene of L. crispatus strain F5.7 (lbs gene). A sequence encoding the green fluorescent protein (GFP) was inserted as reporter gene, and an erythromycin resistance gene was added as selective marker. All constructs were able to express GFP in the cloning host E. coli XL1-Blue and different Lactobacillus strains as verified by FACS and laser scanning confocal microscopy. Conclusion Lactobacillus isolated from gastrointestinal tract of broiler chickens and selected for probiotic characteristics can be genetically modified by introducing an expression cassette into the lbs locus. The transformed bacteria expressed on its cell wall surface different fluorescent proteins used as reporters of promoter function. It is possible then that similar bacterial model expressing pathogen antigens can

  18. Development of a Markerless Genetic Exchange System in Desulfovibrio vulgaris Hildenborough and Its Use in Generating a Strain with Increased Transformation Efficiency

    Energy Technology Data Exchange (ETDEWEB)

    Keller, Kimberly L.; Bender, Kelly S.; Wall, Judy D.

    2009-07-21

    In recent years, the genetic manipulation of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough has seen enormous progress. In spite of this progress, the current marker exchange deletion method does not allow for easy selection of multiple sequential gene deletions in a single strain because of the limited number of selectable markers available in D. vulgaris. To broaden the repertoire of genetic tools for manipulation, an in-frame, markerless deletion system has been developed. The counterselectable marker that makes this deletion system possible is the pyrimidine salvage enzyme, uracil phosphoribosyltransferase, encoded by upp. In wild-type D. vulgaris, growth was shown to be inhibited by the toxic pyrimidine analog 5-fluorouracil (5-FU); whereas, a mutant bearing a deletion of the upp gene was resistant to 5-FU. When a plasmid containing the wild-type upp gene expressed constitutively from the aph(3')-II promoter (promoter for the kanamycin resistance gene in Tn5) was introduced into the upp deletion strain, sensitivity to 5-FU was restored. This observation allowed us to develop a two-step integration and excision strategy for the deletion of genes of interest. Since this inframe deletion strategy does not retain an antibiotic cassette, multiple deletions can be generated in a single strain without the accumulation of genes conferring antibiotic resistances. We used this strategy to generate a deletion strain lacking the endonuclease (hsdR, DVU1703) of a type I restriction-modification system, that we designated JW7035. The transformation efficiency of the JW7035 strain was found to be 100 to 1000 times greater than that of the wild-type strain when stable plasmids were introduced via electroporation.

  19. Micro-shock Wave Assisted Plant Transformation

    Science.gov (United States)

    Gnanadhas, Divya Prakash; Datey, Akshay; Chakravortty, Dipshikha; Gopalan, Jagadeesh

    Genetically modified (GM) crops are developed by transforming the desired DNA to plant. There are various methods employed to achieve the required transformation in plants. Agrobacterium mediated transformation and Biolistics or particle bombardment method are the most commonly used methods.

  20. A Two-Step Strategy for System Identification of Civil Structures for Structural Health Monitoring Using Wavelet Transform and Genetic Algorithms

    Directory of Open Access Journals (Sweden)

    Carlos Andres Perez-Ramirez

    2017-01-01

    Full Text Available Nowadays, the accurate identification of natural frequencies and damping ratios play an important role in smart civil engineering, since they can be used for seismic design, vibration control, and condition assessment, among others. To achieve it in practical way, it is required to instrument the structure and apply techniques which are able to deal with noise-corrupted and non-linear signals, as they are common features in real-life civil structures. In this article, a two-step strategy is proposed for performing accurate modal parameters identification in an automated manner. In the first step, it is obtained and decomposed the measured signals using the natural excitation technique and the synchrosqueezed wavelet transform, respectively. Then, the second step estimates the modal parameters by solving an optimization problem employing a genetic algorithm-based approach, where the micropopulation concept is used to improve the speed convergence as well as the accuracy of the estimated values. The accuracy and effectiveness of the proposal are tested using both the simulated response of a benchmark structure and the measurements of a real eight-story building. The obtained results show that the proposed strategy can estimate the modal parameters accurately, indicating than the proposal can be considered as an alternative to perform the abovementioned task.

  1. 应用正交试验法优选茎瘤芥高效遗传转化体系%Optimization of Genetic Transformation System of Tumorous Stem Mastard by Orthogonal Design

    Institute of Scientific and Technical Information of China (English)

    梁婷; 刘毅; 蔡应繁; 岳显可; 江怀仲; 何晓红

    2012-01-01

    [ Objective ] To establish the genetic transformation system, so us to lay foundation lor the genetic transformation of tumoroua stem mustard and other Brassica plants. [ Method] The effects of different time of precullure.cocullure,bacteria liquid concentration and dipping time on the transformation rate of lurnorous stem mustard were studied with orthogonal design. [ Result ] The best conditions for genetic transformation were 3 d preculture,4 d coculture, bacteria liquid concentration (OD600) 0.5 and 10 min dipping,then the Kan concentration was 30 mg/L. [Conclution] The optimum conditions for the transformation of tumorous stem mustard were selected, which greatly improved the transformation rale.%[目的]建立茎瘤芥高效遗传转化体系,为茎瘤芥及芸薹属植物遗传转化奠定基础.[方法]应用正交试验法研究茎瘤芥遗传转化过程中预培养时间、共培养时间、侵染液浓度、侵染时间、对转化率的影响.[结果]最适合茎瘤芥CRY2基因RNAi载体遗传转化的条件为预培养3d、共培养4d、侵染液浓度OD600=0.5、侵染10 min,此时Kana选择压为30mg/L.[结论]优选出了农杆菌转化的最宜条件,遗传转化率有较大提高.

  2. Model Transformations? Transformation Models!

    NARCIS (Netherlands)

    Bézivin, J.; Büttner, F.; Gogolla, M.; Jouault, F.; Kurtev, I.; Lindow, A.

    2006-01-01

    Much of the current work on model transformations seems essentially operational and executable in nature. Executable descriptions are necessary from the point of view of implementation. But from a conceptual point of view, transformations can also be viewed as descriptive models by stating only the

  3. Chickpea regeneration and genetic transformation

    African Journals Online (AJOL)

    STORAGESEVER

    2010-02-08

    Feb 8, 2010 ... embryo. Grafting. Establishmen t of plants. Chakraborti et al., 2006. C-235. MS. TDZ, 2-iP ..... Plant protoplasts (single cell system) culture has been exciting ..... 15 days old leaf and stem explants with wild as well as disarmed ...

  4. Progress in planta transformation without tissue culture

    Institute of Scientific and Technical Information of China (English)

    GU Yun-Hong; YU Zeng-Liang; QIN Guang-Yong; HUO Yu-Ping

    2004-01-01

    With the development of planta genetic engineering, more emphases have been laid on convenient and high efficient genetic transformation methods. And transformation without tissue culture is a prospective direction of it. In this paper, traditional transformation methods and the methods of non-tissue culture were summarized. With the exploration and application of Arabidopsis transformation mechanism, with the use of ion beam-mediated transformation invented by Chinese scientists and the development of other transformation methods, transformation methods without tissue culture and planta genetic engineering could be improved rapidly.

  5. Research Progress on the Influencing Factors of Agrobacterium-mediated Genetic Transformation of Cannabis sativa L.%农杆菌介导的汉麻遗传转化影响因素的研究进展

    Institute of Scientific and Technical Information of China (English)

    赵越; 魏国江; 潘冬梅; 韩承伟; 韩喜财; 徐磊

    2015-01-01

    Genetic transformation technology is the basis to develop new hemp varieties and study functional ge‐nomics .The efficiency of A grobacterium‐mediated Cannabis sativa L .genetic transformation was effected by genotype ,explant size ,agrobacterium strains ,pretreatment time ,infection time ,co‐culture time ,antioxidants ,a‐cetyl clove ketone and so on .These influencing factors and the research progress were analyzed and summa‐rized ,so as to provide the scientific basis for promoting Cannabis sativa L .genetic transformation research .%遗传转化技术是培育汉麻新品种和进行功能基因组学研究的基础,采用农杆菌介导法对汉麻进行遗传转化,汉麻的基因型、外植体大小、农杆菌菌株、预处理时间、侵染时间、共培养时间、抗氧化剂、乙酰丁香酮等都会对遗传转化效率产生影响。分析并概括了这些影响因素及其研究进展情况,为促进汉麻农杆菌介导的遗传转化研究提供科学依据。

  6. Hardware Implementation of a Genetic Algorithm Based Canonical Singed Digit Multiplierless Fast Fourier Transform Processor for Multiband Orthogonal Frequency Division Multiplexing Ultra Wideband Applications

    Directory of Open Access Journals (Sweden)

    Mahmud Benhamid

    2009-01-01

    Full Text Available Problem statement: Ultra Wide Band (UWB technology has attracted many researchers' attention due to its advantages and its great potential for future applications. The physical layer standard of Multi-band Orthogonal Frequency Division Multiplexing (MB-OFDM UWB system is defined by ECMA International. In this standard, the data sampling rate from the analog-to-digital converter to the physical layer is up to 528 M sample sec-1. Therefore, it is a challenge to realize the physical layer especially the components with high computational complexity in Very Large Scale Integration (VLSI implementation. Fast Fourier Transform (FFT block which plays an important role in MB-OFDM system is one of these components. Furthermore, the execution time of this module is only 312.5 ns. Therefore, if employing the traditional approach, high power consumption and hardware cost of the processor will be needed to meet the strict specifications of the UWB system. The objective of this study was to design an Application Specific Integrated Circuit (ASIC FFT processor for this system. The specification was defined from the system analysis and literature research. Approach: Based on the algorithm and architecture analysis, a novel Genetic Algorithm (GA based Canonical Signed Digit (CSD Multiplier less 128-point FFT processor and its inverse (IFFT for MB-OFDM UWB systems had been proposed. The proposed pipelined architecture was based on the modified Radix-22 algorithm that had same number of multipliers as that of the conventional Radix-22. However, the multiplication complexity and the ROM memory needed for storing twiddle factors coefficients could be eliminated by replacing the conventional complex multipliers with a newly proposed GA optimized CSD constant multipliers. The design had been coded in Verilog HDL and targeted Xilinx Virtex-II FPGA series. It was fully implemented and tested on real hardware using Virtex-II FG456 prototype board and logic analyzer

  7. A mutation in negative regulator of basal resistance WRKY17 of Arabidopsis increases susceptibility to Agrobacterium-mediated transient genetic transformation [v2; ref status: indexed, http://f1000r.es/ya

    Directory of Open Access Journals (Sweden)

    Benoît Lacroix

    2013-02-01

    Full Text Available Agrobacterium is a phytopathogenic bacterium that induces crown gall disease in many plant species by transferring and integrating a segment of its own DNA (T-DNA into its host genome. Whereas Agrobacterium usually does not trigger an extensive defense response in its host plants, it induces the expression of several defense-related genes and activates plant stress reactions. In the complex interplay between Agrobacterium and its host plant, Agrobacterium has evolved to take advantage of these plant defense pathways for its own purpose of advancement of the infection process. For example, Agrobacterium utilizes the host stress response transcriptional regulator VIP1 to facilitate nuclear import and proteasomal uncoating of its T-DNA during genetic transformation of the host cell. In Arabidopsis, the VIP1 gene expression is repressed by WRKY17, a negative regulator of basal resistance to Pseudomonas. Thus, we examined whether WRKY17 is also involved in plant susceptibility to genetic transformation by Agrobacterium. Using reverse genetics, we showed that a wrky17 mutant displays higher expression of the VIP1 gene in roots, but not in shoots. In a root infection assay, the wrky17 mutant plants were hyper-susceptible to Agrobacterium compared to wild type plants. WRKY17, therefore, may act as a positive regulator of Arabidopsis resistance to Agrobacterium. This notion is important for understanding the complex regulation of Agrobacterium-mediated transient genetic transformation; thus, although this paper reports a relatively small set of data that we do not plan to pursue further in our lab, we believe it might be useful for the broad community of plant pathologists and plant biotechnologists.

  8. A mutation in negative regulator of basal resistance WRKY17 of Arabidopsis increases susceptibility to Agrobacterium-mediated genetic transformation [v1; ref status: indexed, http://f1000r.es/no

    Directory of Open Access Journals (Sweden)

    Benoît Lacroix

    2013-02-01

    Full Text Available Agrobacterium is a phytopathogenic bacterium that induces crown gall disease in many plant species by transferring and integrating a segment of its own DNA (T-DNA into its host genome. Whereas Agrobacterium usually does not trigger an extensive defense response in its host plants, it induces the expression of several defense-related genes and activates plant stress reactions. In the complex interplay between Agrobacterium and its host plant, Agrobacterium has evolved to take advantage of these plant defense pathways for its own purpose of advancement of the infection process. For example, Agrobacterium utilizes the host stress response transcriptional regulator VIP1 to facilitate nuclear import and proteasomal uncoating of its T-DNA during genetic transformation of the host cell. In Arabidopsis, the VIP1 gene expression is repressed by WRKY17, a negative regulator of basal resistance to Pseudomonas. Thus, we examined whether WRKY17 is also involved in plant susceptibility to genetic transformation by Agrobacterium. Using reverse genetics, we showed that a wrky17 mutant displays higher expression of the VIP1 gene in roots, but not in shoots. In a root infection assay, the wrky17 mutant plants were hyper-susceptible to Agrobacterium compared to wild type plants. WRKY17, therefore, may act as a positive regulator of Arabidopsis resistance to Agrobacterium. This notion is important for understanding the complex regulation of Agrobacterium-mediated genetic transformation; thus, although this paper reports a relatively small set of data that we do not plan to pursue further in our lab, we believe it might be useful for the broad community of plant pathologists and plant biotechnologists.

  9. A novel gateway-compatible binary vector series (PC-GW) for flexible cloning of multiple genes for genetic transformation of plants.

    Science.gov (United States)

    Dalal, Jyoti; Yalamanchili, Roopa; La Hovary, Christophe; Ji, Mikyoung; Rodriguez-Welsh, Maria; Aslett, Denise; Ganapathy, Sowmya; Grunden, Amy; Sederoff, Heike; Qu, Rongda

    2015-09-01

    The rapidly advancing field of plant synthetic biology requires transforming plants with multiple genes. This has sparked a growing interest in flexible plant transformation vectors, which can be used for multi-gene transformations. We have developed a novel binary vector series, named the PC-GW series (GenBank: KP826769-KP826773), for Agrobacterium-mediated plant transformation. The PC-GW vectors use the pCAMBIA vector backbone, and contain NPTII, hpt, bar, mCherry or egfp genes as selectable markers for plant transformation. In a modified multiple cloning site (MCS) of the T-DNA region, we have placed the attR1, attR2 and ccdB sequences for rapid cloning of one to four genes by Gateway™-assisted recombination. In addition, we have introduced four meganuclease sites, and other restriction sites for multi-gene vector construction. Finally, we have placed a CaMV 35S promoter and a 35S terminator on the 5' and 3' ends of the MCS. The CaMV 35S promoter is flanked by PstI restriction sites that can be used to replace it with another promoter sequence if needed. The PC-GW vectors provide choices for selectable markers, cloning methods, and can accommodate up to eight gene constructs in a single T-DNA, thereby significantly reducing the number of transformations or crosses needed to generate multi-transgene expressing plants.

  10. Agrobacterium-mediated transformation: rice transformation.

    Science.gov (United States)

    Slamet-Loedin, Inez H; Chadha-Mohanty, Prabhjit; Torrizo, Lina

    2014-01-01

    Agrobacterium is a common soil bacterium with natural capacity for trans-kingdom transfer of genetic information by transferring its T-DNA into the eukaryotic genome. In agricultural plant biotechnology, combination of non-phytopathogenic strain of Agrobacterium tumefaciens with modified T-DNA and vir-genes in a binary vector system is the most widely utilized system for genetic improvement in diverse plant species and for gene function validation. Here we have described a highly efficient A. tumefaciens-mediated transformation system for indica and japonica rice cultivars based on an immature embryo system.

  11. Restriction enzyme improves the efficiency of genetic transformations in Moniliophthora perniciosa, the causal agent of witches’ broom disease in Theobroma cacao

    Directory of Open Access Journals (Sweden)

    Francis Julio Fagundes Lopes

    2008-02-01

    Full Text Available The presence of restriction enzymes in the transformation mixture improved the efficiency of transformation in Moniliophthora perniciosa. The influence of the vector shape (linear or circular, the patterns of plasmid integration in genomic sites and the influence of the promoter used to express the gene marker were also analyzed. The addition of BamHI or NotI increased the number of transformants by 3-10-fold and 3-fold, respectively, over the control without added enzyme. The use of pre-linearized plasmid did not increase the transformation efficiency in comparison with the circular plasmid. However, the frequency of multi-copy transformants increased significantly. The transformation procedure here reported resulted in better production of protoplasts and transformation efficiency. In addition, the time necessary for the detection of the first transformants and the number of insertions were reduced.A presença de enzima de restrição na mistura de transformação aumentou a eficiência da transformação em Moniliophthora perniciosa. A influência da forma do vetor (linear ou circular, o padrão de integração do plasmídeo nos sítios genômicos e a influência do promotor usado para expressar o gene marcador foram também analisados. A adição de BamHI ou NotI aumentou o número de transformantes 3-10 vezes e 3 vezes, respectivamente, em relação ao controle sem a adição da enzima. O uso de plasmídeos pré-linearizados não aumentou a eficiência da transformação quando comparado à eficiência obtida com plasmídeos circulares. No entanto, a freqüência de transformantes multi-cópias aumentou significativamente. Juntos os procedimentos reportados aqui resultaram em processos mais eficientes de produção de protoplastos e transformação, onde o tempo necessário para o aparecimento dos transformantes e o número de inserções múltiplas foi reduzido.

  12. Polyethylene glycol (PEG)-mediated transformation in filamentous fungal pathogens

    Science.gov (United States)

    Genetic transformation is an essential tool in molecular biology for many purposes including the study of gene function and the genetic improvement of an organism. The genetic transformation of many fungal species is a well established process that can be carried out by utilizing different transform...

  13. A novel homologous dominant selection marker for genetic transformation of Penicillium chrysogenum: overexpression of squalene epoxidase-encoding ergA.

    Science.gov (United States)

    Sigl, Claudia; Handler, Monika; Sprenger, Georg; Kürnsteiner, Hubert; Zadra, Ivo

    2010-11-01

    Genetic engineering requires genetic selection markers. For generation of biosafe strains in industrial applications, homologous dominant selection markers allowing "self-cloning" are best suited but scarce. Here we describe a novel homologous dominant genetic selection system for the filamentous fungus Penicillium chrysogenum based on overexpression of the P. chrysogenum squalene epoxidase-encoding ergA gene, which confers resistance against terbinafine. Terbinafine (TRB) is a potent antifungal drug used in therapy of fungal infections. Overexpression of ergA was driven by the P. chrysogenum endoxylanase xylP promoter that is highly inducible by xylose. The suitability of the novel selection marker cassette for genetic manipulation was proven by its use for targeted deletion of the transcription factor nosA in P. chrysogenum. NosA-deficiency did not affect growth rates on solid or in liquid media, conidiation in light or darkness, and resistance to hydrogen peroxide. However, NosA-deficiency significantly decreased penicillin productivity. As TRB inhibits the growth of a variety of fungal species, this novel selection marker is expected to be suitable for genetic engineering of diverse fungal species.

  14. 基于量子遗传改进支持向量机理论的变压器故障诊断%The Fault Diagnosis of Transformers with Support Vector Machine Theory Improved by Quantum Genetic Algorithm

    Institute of Scientific and Technical Information of China (English)

    于虹; 孙鹏

    2011-01-01

    提出了基于量子遗传改进支持向量机理论的变压器故障诊断方法.该算法首先利用粗糙集技术时变压器知识进行属性约简,通过属性表获得变压器故障的最简决策表以作为支持向量机的输入,并利用量子遗传算法获得支持向量机的最优参数设置.实验结果表明,该诊断方法分类性能良好、可靠性高且有效可行.%A new fault diagnosis method is proposed for transformers, which is based on support vector machine theory improved by quantum genetic algorithm. Firstly. The algorithm is as follows: to do attribute reduction to the transformers knowledge with the technologies of rough set, to get the simple decision table for faults and imput the table into the support vector machine, and to achieve the optimum parameters setting of the support vector machine with the quantum genetic algorithm. The experimental results demonstrate that the proposed method works well in the fault classification and is reliable, effective and feasible.

  15. Genetic Transformation of Sweet Potato and Its Application in Molecular Breeding%甘薯遗传转化及其在分子育种中的应用

    Institute of Scientific and Technical Information of China (English)

    杨俊; 张敏; 张鹏

    2011-01-01

    甘薯是全球重要的块根类作物,不仅为人类提供了粮食,同时也是动物饲料、淀粉及燃料乙醇等工业加工的原材料.建立甘薯遗传转化体系不仅是开展甘薯重要基因功能验证的平台,同时也是甘薯转基因分子育种的必要手段.本文对甘薯遗传转化技术的发展及应用进行了综述.%Sweet potato is an important root crop for food production world-wide and serves as raw material for the processing of feeds, starches and bioethanol in various industries. Establishment of efficient genetic transformation system in sweet potato not only provides a platform for functional verification of important genes, but also is an essential tool for molecular breeding in sweet potato improvement. This review presents the development and application of genetic transformation in sweet potato.

  16. The effect of the number of observations used for Fourier transform infrared model calibration for bovine milk fat composition on the estimated genetic parameters of the predicted data

    NARCIS (Netherlands)

    Rutten, M.J.M.; Bovenhuis, H.; Arendonk, van J.A.M.

    2010-01-01

    Fourier transform infrared spectroscopy is a suitable method to determine bovine milk fat composition. However, the determination of fat composition by gas chromatography, required for calibration of the infrared prediction model, is expensive and labor intensive. It has recently been shown that the

  17. Establishment of Agrobacterium tumefaciens-mediated Genetic Transformation System for Aspergillus awamori%根癌农杆菌介导的泡盛曲霉遗传转化系统的构建

    Institute of Scientific and Technical Information of China (English)

    陈凤; 王坤; 银超; 李德明; 任楠; 李俊星

    2012-01-01

    [Objective] This study aimed to establish Agrobaclerium tumefaciens-me.d'ialed genetic transformation system forAspergillus awamori and investigate the feasibility of expressing helerologous proteins in A. awamori. [ Method] Appropriate A. awamori host strains were deter-mined according to the secretory protein profile. Selectable marker was selected for genetic transformation by drug sensitivity analysis. The es-tablished A. awamori genetic transformation system was used for transformation and expression analysis of Rhizomucor miehei lipase ( RML). The feasibility of using A. awamori to express heleroJogous proteins was investigated by identification of transfbrmants and property analysis, [ Result ] Based on the analysis of secretory protein profile, A. awamori strains CBSl 15. 52 and CICC2257 were determined as the host .strains for helerologous protein expression; drug sensitivity analysis shows that hygromycin B resistance gene (HygBr) is an effective genetic select-able marker; by using Agrobactenum tumefaciens-mediated transformation ( ATMT) method, the plasmid pHGW-amdS containing HygBr was successfully transformed into A. awamori strain CBSl 15. 52 to establish the genetic transformation system of A. awamori with HygB' as select-able marker. RML was transformed into A. awamon and its expression was validated by substrate hydrolysis test, SDS-PAGE and Western blot. [ Conclusion ] This .study demonstrates that the genetic transformation system of A. awamori mediated by Agrobaclerium tumefaciens has potential feasibility for expression of heterologous proteins.%[目的]建立根癌农杆菌介导的泡盛曲霉遗传转化系统,探讨利用泡盛曲霉表达异源蛋白的可行性.[方法]从国内主要菌种库中购买泡盛曲霉菌株,根据分泌蛋白图谱分析确定合适的宿主菌,在此基础上通过药物敏感性分析确定遗传转化的选择标记,并利用构建的泡盛曲霉遗传转化体系对米黑根毛霉脂肪酶基因RML进

  18. Genetic transformation of European chestnut somatic embryos with a native thaumatin-like protein (CsTL1) gene isolated from Castanea sativa seeds.

    Science.gov (United States)

    Corredoira, Elena; Valladares, Silvia; Allona, Isabel; Aragoncillo, Cipriano; Vieitez, Ana M; Ballester, Antonio

    2012-11-01

    The availability of a system for direct transfer of antifungal candidate genes into European chestnut (Castanea sativa Mill.) would offer an alternative approach to conventional breeding for production of chestnut trees tolerant to ink disease caused by Phytophthora spp. For the first time, a chestnut thaumatin-like protein gene (CsTL1), isolated from chestnut cotyledons, has been overexpressed in three chestnut somatic embryogenic lines. Transformation experiments have been performed using an Agrobacterium tumefaciens Smith and Townsend vector harboring the neomycin phosphotransferase (NPTII) selectable and the green fluorescent protein (EGFP) reporter genes. The transformation efficiency, determined on the basis of the fluorescence of surviving explants, was clearly genotype dependent and ranged from 32.5% in the CI-9 line to 7.1% in the CI-3 line. A total of 126 independent transformed lines were obtained. The presence and integration of chestnut CsTL1 in genomic DNA was confirmed by polymerase chain reaction (PCR) and Southern blot analyses. Quantitative real-time PCR revealed that CsTL1 expression was up to 13.5-fold higher in a transgenic line compared with its corresponding untransformed line. In only one of the 11 transformed lines tested, expression of the CsTL1 was lower than the control. The remaining 115 transformed lines were successfully subjected to cryopreservation. Embryo proliferation was achieved in all of the transgenic lines regenerated and the transformed lines showed a higher mean number of cotyledonary stage embryos and total number of embryos per embryo clump than their corresponding untransformed lines. Transgenic plants were regenerated after maturation and germination of transformed somatic embryos. Furthermore, due to the low plantlet conversion achieved, axillary shoot proliferation cultures were established from partially germinated embryos (only shoot development), which were multiplied and rooted according to procedures already

  19. 豇豆子叶节再生体系建立及Pti4基因的遗传转化%Establishment of Regeneration System of Cotyledon Node and Genetic Transformation ofPti4 Gene to Cowpea

    Institute of Scientific and Technical Information of China (English)

    姜建业; 谢永东; 王一鸣; 赵丽; 铁曼曼; 唐懿

    2015-01-01

    为获得豇豆(Vigna unguiculata)遗传转化体系,以‘成豇七号’带1片子叶的子叶节作为外植体,对其高效再生体系和农杆菌介导抗病基因Pti4的遗传转化进行了研究。结果表明,豇豆无菌苗、不定芽诱导和不定芽伸长培养的最适培养基分别为MSB5+6-BA 3.0 mg L–1、MSB5+6-BA 1.0 mg L–1+ KT 0.06 mg L–1和MSB5+6-BA 0.5 mg L–1+ IBA 0.2 mg L–1。不定芽在MS培养基上能迅速诱导生根,获得完整植株。以豇豆子叶节为受体,通过农杆菌介导成功将Pti4整合到‘成豇七号’抗性芽基因组中。因此,豇豆高效再生体系的建立为遗传育种研究奠定了基础。%In order to obtain genetic transformation system of cowpea (Vigna unguiculata), its cotyledon nodes with one cotyledon of ‘Chengjiang 7’ was used as explants, the regeneration system andPti4 genetic transformation system byAgrobacterium-medium were studied. The results showed that the optimum mediums for aseptic seedling, adventitious bud introduction and adventitious bud elongation were MSB5 + 6-BA 3.0 mg L–1, MSB5 + 6-BA 1.0 mg L–1 + KT 0.06 mg L–1 and MSB5 + 6-BA 0.5 mg L–1 + IBA 0.2 mg L–1, respectively. Adventitious buds could rapidly rooting on MS medium to obtain full plantlets. The cotyledon node was used as receptor, thepit4 gene mediated byAgrobacterium was successfully transformed into the genome of resistant buds of ‘Chengjiang 7’. Therefore, the establishment of high efifcient regeneration system for cowpea laid basis for studying on genetic transformation.

  20. Genetic transformation of Indian bread (T. aestivum) and pasta (T. durum) wheat by particle bombardment of mature embryo-derived calli

    OpenAIRE

    Khurana Paramjit; Patnaik Debasis

    2003-01-01

    Abstract Background Particle bombardment has been successfully employed for obtaining transgenics in cereals in general and wheat in particular. Most of these procedures employ immature embryos which are not available throughout the year. The present investigation utilizes mature seeds as the starting material and the calli raised from the hexaploid Triticum aestivum and tetraploid Triticum durum display a high regeneration response and were therefore used as the target tissue for genetic tra...

  1. Efficient genetic transformation of Lotus corniculatus L. using a direct shoot regeneration protocol, stepwise hygromycin B selection, and a super-binary Agrobacterium tumefaciens vector

    Directory of Open Access Journals (Sweden)

    Nikolić Radomirka

    2007-01-01

    Full Text Available Cotyledons from 6-day-old Lotus corniculatus cv. Bokor seedlings, transversally cut into two halves, were capa­ble of regenerating buds without intervening callus formation. The explants were co-cultivated with the Agrobacterium tumefaciens LBA4404/pTOK233 superbinary vector carrying the uidA-intron gene and the genes hpt and nptII. They were cultured for 14 days on a regeneration medium, then subjected to a stepwise hygromycin B selection procedure consisting of gradually increasing antibiotic concentrations (5-15 mg L-1 over 21 weeks. Transformed shoots were obtained within 5 months after co-cultivation. Out of 124 initially co-cultivated explants, 52 (42% plants survived hygromycin B selection. The presence of transgenes in regenerated plants was verified by β-glucuronidase histochemical assays and PCR analysis for the presence of uidA gene sequences. Hygromycin B-resistant and PCR-positive T0 plants were cultured in the greenhouse to produce flowers and seeds. The obtained data demonstrate that the reported transformation protocol could be useful for introducing agriculturally important genes into the new L. corniculatus cultivar Bokor.

  2. Molecular Cloning of a Cellulose Synthase Gene PtoCesA1 from Populus tomentosa and Its Genetic Transformation in Tobacco

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    A 3 125 bp cellulose synthase gene, PtoCesA1, which has a 98% identity to PtrCesA1 from Populus tremuloides,was cloned from cDNA prepared from secondary xylem of P. tomentosa. Four anti-expression vectors with different fragments of PtoCesA1, named as pBIPF, pBICC1, pBIPR and pBIBR, were constructed. Some traits of transformed tobacco of pBICC1, pBIPR and pBIBR differed from wild types, such as small leaves, "dwarf" phenotype and thinner xylem and fiber cell walls than wild plants consistent with a loss of cellulose. It indicated that the growth of transgenic tobacco was restrained by the expression of anti-PtoCesA1. Transgenic tobacco was obtained and the contents of cellulose and lignin were analyzed as well as the width and length of fiber cells, and xylem thickness for both transgenic and control plants. Transformed tobacco showed a different phenotype from control plants and it implied that PtoCesA1 was essential for the cellulose biosynthesis in poplar stems.

  3. 改进遗传算法在干式电力变压器优化设计中的应用%Application of Improved Genetic Algorithm in Optimization Design of Dry Type Power Transformer

    Institute of Scientific and Technical Information of China (English)

    韩能霞

    2014-01-01

    Introduction was made to the basic principle and algorithm steps of genetic algorithm. This paper rebuilt its coding and decoding plans, restricting conditions’ treatment, searching speed and global optimization etc., so as to enable new mass body’s quality better and to quicken the optimization process. The improved genetic algorithm was adopted to carry out optimization for electromagnetic calculation de-sign of dry type power transformer, applied in the single goal and dual goal optimization design for SGB-800/10 dry type power transformer. The actual example parameters comparison and analysis show that the algorithm is actually feasible with better optimization effect.%介绍了遗传算法的基本原理及算法步骤,对其编码与解码方案、约束条件的处理、搜索速度、全局寻优等进行了改造,使新群体的整体素质更好,加快了寻优过程。采用改进的遗传算法对干式变压器的电磁计算设计进行了优化,并应用于SGB-800/10干式电力变压器的单目标和双目标的优化设计。通过实例参数对比分析,表明该算法切实可行,优化效果较好。

  4. Developing a survey instrument to assess the readiness of primary care data, genetic and disease registries to conduct linked research: TRANSFoRm International Research Readiness (TIRRE survey instrument

    Directory of Open Access Journals (Sweden)

    Emily Leppenwell

    2013-05-01

    Full Text Available Background Clinical data are collected for routine care in family practice; there are also a growing number of genetic and cancer registry data repositories. The Translational Research and Patient Safety in Europe (TRANSFoRm project seeks to facilitate research using linked data from more than one source. We performed a requirements analysis which identified a wide range of data and business process requirements that need to be met before linking primary care and either genetic or disease registry data.Objectives To develop a survey to assess the readiness of data repositories to participate in linked research – the Transform International Research Readiness (TIRRE survey.Method We develop the questionnaire based on our requirement analysis; with questions at micro-, meso- and macro levels of granularity, study-specific questions about diabetes and gastro-oesophageal reflux disease (GORD, and research track record. The scope of the data required was extensive. We piloted this instrument, conducting ten preliminary telephone interviews to evaluate the response to the questionnaire.Results Using feedback gained from these interviews we revised the questionnaire; clarifying questions that were difficult to answer and utilising skip logic to create different series of questions for the various types of data repository. We simplified the questionnaire replacing free-text responses with yes/no or picking list options, wherever possible. We placed the final questionnaire online and encouraged its use (www.clininf.eu/jointirre/info.html.Conclusion Limited field testing suggests that TIRRE is capable of collecting comprehensive and relevant data about the suitability and readiness of data repositories to participate in linked data research.

  5. Ti和Ri质粒对栝楼双转化的研究%Genetic Transformation of Hairy Roots in Trichosanthes kirilowii Maxim.by Ti and Ri Plasmids

    Institute of Scientific and Technical Information of China (English)

    雷和田; 宋经元; 祁荫麟; 张荫麟; 杨峻山; 郭志刚

    2001-01-01

    目的:建立栝楼双转化毛状根培养系统。方法:①利用根癌农杆菌Agrobacterium tumefaciens菌株C58感染栝楼Trichosanthes kirilowii.无菌苗诱导冠瘿瘤,然后用发根农杆菌Agrobacterium rhizogenes 菌株15834感染其再生植株,诱导出毛状根;②用纸电泳检测Ti和Ri质粒是否转化成功;③运用分光光度计和SDS-PAGE检测栝楼组织中所含蛋白。结果:Ti和Ri质粒转化成功并建立了栝楼双转化毛状根培养系统。结论:双转化毛状根与一般毛状根相比,具有生长更迅速等特点而蛋白含量基本一样;因此,利用它来生产天花粉蛋白更具有开发潜力。%Objective:To estabish a hairy root culture system by doubletransfonmation for Trichosathes kirilowii. Method:①Crown galls were induced by direct infection of sterile seedlings of T.kirilonii with Agrobacterium tumefaciens C58, and then the hairy roots were obtained from the regenerated plants by infection with A.rhzogenes 15834;②Transformation of Ti and Ri plasmids was inspected by high-pressure-paper electrophoresis;③ The protein contents in the tissues of T.kirilowii were inspected by spectrophotometer and SDS-PAGE.Result:A hairy root culture system has been established sucessfully by double transformation with Ti and Ri plasmids in T. kirilowii.Conclusion:Compared with the ordinary hairy roots, the double transformed hairy roots grow faster but retain similar protein contents.

  6. Genetic Transformation of Tobacco with the Trehalose Synthase Gene from Grifola frondosa Fr. Enhances the Resistance to Drought and Salt in Tobacco

    Institute of Scientific and Technical Information of China (English)

    Shu-Zhen ZHANG; Ben-Peng YANG; Cui-Lian FENG; Huo-Long TANG

    2005-01-01

    Trehalose is a non-reducing disaccharide of glucose that functions as a protectant in the stabilization of biological structures and enhances the tolerance of organisms to abiotic stress. In the present study, we report on the expression of the Grifolafrondosa Fr. trehalose synthase (TSase) gene for manipulating abiotic stress tolerance in tobacco (Nicotiana tabaccum L.). The expression of the transgene was under the control of two tandem copies of the CaMV35S promoter and was transferred into tobacco by Agrobacterium tumefaciens EHA105. Compared with non-transgenic plants, transgenic plants were able to accumulate high levels of products of trehalose, which were increased up to 2.126-2.556 mg/g FW, although levels were undetectable in non-transgenic plants. This level of trehalose in transgenic plants was 400-fold higher than that of transgenic tobacco plants cotransformed with Escherichia coli TPS and TPP on independent expression cassettes, twofold higher than that of transgenic rice plants transformed with a bifunctional fusion gene (TPSP) of the trehalose-6-phosphate (T-6-P) synthase (TPS) and T-6-P phosphatase (TPP) of E. coli, and 12-fold higher than that of transgenic tobacco plants transformed the yeast TPS1 gene.It has been reported that transgenic plants with E. coli TPS and/or TPP were severely stunted and had morphological alterations of their roots. Interestingly, our transgenic plants have obvious morphological changes, including thick and deep-coloured leaves, but show no growth inhibition; moreover, these morphological changes can restore to normal type in T2 progenies. Trehalose accumulation in 35S-35S:TSase plants resulted in increased tolerance to drought and salt, as shown by the results of tests on drought, salt tolerance, and drought physiological indices, such as water content in excised leaves, malondialdehyde content, chlorophyll a and b contents, and the activity of superoxide dismutase and peroxidase in excised leaves. These results

  7. Genetic transformation of cotton with a harpin-encoding gene hpaXoo confers an enhanced defense response against different pathogens through a priming mechanism

    Directory of Open Access Journals (Sweden)

    Song Congfeng

    2010-04-01

    Full Text Available Abstract Background The soil-borne fungal pathogen Verticillium dahliae Kleb causes Verticillium wilt in a wide range of crops including cotton (Gossypium hirsutum. To date, most upland cotton varieties are susceptible to V. dahliae and the breeding for cotton varieties with the resistance to Verticillium wilt has not been successful. Results Hpa1Xoo is a harpin protein from Xanthomonas oryzae pv. oryzae which induces the hypersensitive cell death in plants. When hpa1Xoo was transformed into the susceptible cotton line Z35 through Agrobacterium-mediated transformation, the transgenic cotton line (T-34 with an improved resistance to Verticillium dahliae was obtained. Cells of the transgenic T-34, when mixed with the conidia suspension of V. dahliae, had a higher tolerance to V. dahliae compared to cells of untransformed Z35. Cells of T-34 were more viable 12 h after mixing with V. dahliae conidia suspension. Immunocytological analysis showed that Hpa1Xoo, expressed in T-34, accumulated as clustered particles along the cell walls of T-34. In response to the infection caused by V. dahliae, the microscopic cell death and the generation of reactive oxygen intermediates were observed in leaves of T-34 and these responses were absent in leaves of Z35 inoculated with V. dahliae. Quantitative RT-PCR analysis indicated that five defense-related genes, ghAOX1, hin1, npr1, ghdhg-OMT, and hsr203J, were up-regulated in T-34 inoculated with V. dahliae. The up-regulations of these defense-relate genes were not observed or in a less extent in leaves of Z-35 after the inoculation. Conclusions Hpa1Xoo accumulates along the cell walls of the transgenic T-34, where it triggers the generation of H2O2 as an endogenous elicitor. T-34 is thus in a primed state, ready to protect the host from the pathogen. The results of this study suggest that the transformation of cotton with hpa1Xoo could be an effective approach for the development of cotton varieties with the

  8. PEG介导的苹果腐烂病菌原生质体转化%Development of Genetic Transformation system of Valsa mali of apple mediated by PEG

    Institute of Scientific and Technical Information of China (English)

    高静; 李艳波; 柯希望; 康振生; 黄丽丽

    2011-01-01

    [ Objective ] The genetic transformation of Valsa mali var. Mali was developed by PEG-mediated protoplasts transformation. [Method] It was transformed by PEG-induced fusion of protoplasts. The plasmid pBIG2RHPH2-GFP-GUS carrying hph gene was used and Valsa mali var. Mali 03-8 isolate was used as the host strain. [Result] At 50 mg/mL driselase +10 mg/mL lysing enzymes concentration, the mycelium of Valsa mali var. Mali cultured in YEPD medium for 48 h was hydrolyzed in 10 mL enzymes liquid /0. 5 g wet mycelium for 2 h. The protoplast yield was 4 x 107 CFU/mg. The transformation efficiency was 44 per g DNA. Analysis of the transformants by PCR and Southern blotting showed that the selectable marker gene hph was integrated effectively into the genome of Valsa mali var. Mali. After 5 subculturing on PDA, 87.5% transformants could grow. This stability test of transformants suggested that the foreign gene hph was stable in heredity. [Conclusion] This transformation system is a valuable and important tool for the further study of the pathogenic gene of Valsa mali.%[目的]建立PEG介导的苹果腐烂病菌原生质体遗传转化体系.[方法]本文利用带有hph基因的质粒,以苹果腐烂病菌(Valsa mali var.mali )03-8为受体菌株,通过PEG融合法对其原生体进行转化.[结果]于YEPD内培养48 h的菌丝,在酶解液浓度为50 mg/mL Driselase+10 mg/mL Lysing Enzymes情况下,按10 mL酶液/0.5 g湿菌体比例,酶解2h时可以释放出4×107个/mL原生质体,其转化效率为44个/μg DNA.对转化子的PCR检测和Southern杂交分析表明,hph基因已经整合进苹果树腐烂病菌的基因组中.转化子在PDA培养基中继代5次后,87.5%的转化子仍能正常生长,表明外源基因hph能在苹果树腐烂病菌中稳定遗传.[结论]该转化体系的建立为苹果树腐烂病菌致病相关基因的深入研究奠定了基础.

  9. TRANSFORMACIÓN DE PLANTAS MEDIADA POR AGROBACTERIUM: "INGENIERÍA GENÉTICA NATURAL APLICADA" PLANT TRANSFORMATION MEDIATED BY AGROBACTERIUM: "APPLIED NATURAL GENETIC ENGINEERING"

    Directory of Open Access Journals (Sweden)

    Ana Milena Valderrama Fonseca

    2005-06-01

    Full Text Available Agrobacterium tumefaciens tiene la capacidad de transferir ADN entre reinos diferentes. El impacto de este hallazgo ha tenido grandes aplicaciones en diversos campos de la biología vegetal, agricultura y biotecnología. En este artículo se describen los procesos por los cuales Agrobacterium realiza la transferencia de ADN a la planta, puntualizando en 7 eventos fundamentales para la interacción A. tumefaciens-planta y esta dirigido a profesionales de las áreas biológicas que estén interesados en actualizarse en el tema. El conocimiento básico sobre el mecanismo de transferencia del ADN ha permitido el desarrollo de vectores para la introducción de genes foráneos, dentro de los cuales se describen los 2 tipos de vectores utilizados en la actualidad: co-integrados y binarios. Así mismo se detallan algunos factores importantes que median la transformación y algunas de las principales aplicaciones de la transformación de plantas y hongos mediada por Agrobacterium.Agrobacterium tumefaciens has the ability to transfer DNA between different kingdoms. This finding has had a great impact in several fields of plant biology, agriculture and biotechnology. This review describe the mechanisms by which Agrobacterium transfer DNA to the plant, emphasizing 7 fundamental steps in the A. tumefaciens-plant interaction, to provide an opportunity for professionals in the biological sciences to familiarize themselves with the topic. Basic knowledge about the DNA transfer mechanism has allowed the development of various vectors for the introduction of foreign genes, among which two classes that are currently used are described: co-integrative and binary vectors. Furthermore, detail some important factors that mediate transformation and some of the principal applications of the transformation of plants and fungi by means of Agrobacterium.

  10. Hadamard Transforms

    CERN Document Server

    Agaian, Sos; Egiazarian, Karen; Astola, Jaakko

    2011-01-01

    The Hadamard matrix and Hadamard transform are fundamental problem-solving tools in a wide spectrum of scientific disciplines and technologies, such as communication systems, signal and image processing (signal representation, coding, filtering, recognition, and watermarking), digital logic (Boolean function analysis and synthesis), and fault-tolerant system design. Hadamard Transforms intends to bring together different topics concerning current developments in Hadamard matrices, transforms, and their applications. Each chapter begins with the basics of the theory, progresses to more advanced

  11. Establishment of an Agrobacterium tumefaciens-mediated genetic transformation system of Lilium speciosum%农杆菌介导的药百合鳞片遗传转化体系的建立

    Institute of Scientific and Technical Information of China (English)

    储俊; 许娜; 张强; 王松华; 周正义; 金光明

    2011-01-01

    本试验以药百合鳞片高频再生体系为基础,研究了农杆菌介导的药百合鳞片遗传转化的几个影响因素.结果表明,预培养5d有利于转化;共培养3d并且添加100μmol/L乙酰丁香酮有利于提高转化频率并抑制了农杆菌的过度生长;在侵染阶段添加150 μmol/L乙酰丁香酮,农杆菌侵染液pH值5.7、光吸收值0.8,侵染时间10 min为最佳遗传转化体系.再生植株经gus基因的瞬时表达检测及hpt基因PCR检测证明载体己成功整合到药百合基因组中.%Several factors, affecting Agrobacterium tumefaciens-mediated genetic transformations of lily were studied based on the establishment of regeneration systems. The pre-culture time of 5 days was beneficial for improving transformation frequency. Co-culture for 3 days with 100 μmol/L AS (acetosyringone) resulted not only in higher rates of transformation but also controlled the over-growth of bacteria. The 10-min bacteria infection (ABS600= 0. 8, pH = 5. 7) with 150 μmol/L AS was found to be the optimal selection for transgenic pla-ntlets. It was confirmed by using gus instantaneous expression test and hpt gene PCR test that the vector was integrated into the regeneration genome of lily.

  12. Design on Dual-band Impedance Transformer Based on a Variant of Canonical Genetic Algorithm%基于改进遗传算法的双波带阻抗变换器设计

    Institute of Scientific and Technical Information of China (English)

    史玮; 蔡钧

    2014-01-01

    阻抗匹配技术在微波工程中十分重要。提出了一种基于改进遗传算法的双频带阻抗匹配的设计方法,该阻抗变换器能在2个频率点对任意电阻性负载实现理想的阻抗匹配。通过对多节传输线匹配方程的严格解,得到了双频阻抗变换器的精确设计公式,对用该方法设计的阻抗变换器性能进行了仿真分析,结果显示了该方法的有效性,可用于实际的设计。%The impedance matching technique is very important in microwave engineering. This paper proposes a dual-band impedance matching design method based on a variant of canonical Genetic Algorithm. The impedance transformer can realize ideal im-pedance matching in two frequency points in any resistive load. Based on the exact solution of multi-section transmission line matching equation, the precise design formula of dual band impedance transformer is obtained. The simulation analysis on impedance transforma-tion performances designed by the method is implemented, and the results show that the proposed method is effective and can be used in practical design.

  13. Protocols for In Vitro Propagation, Conservation, Synthetic Seed Production, Embryo Rescue, Microrhizome Production, Molecular Profiling, and Genetic Transformation in Ginger (Zingiber officinale Roscoe.).

    Science.gov (United States)

    Nirmal Babu, K; Samsudeen, K; Divakaran, Minoo; Pillai, Geetha S; Sumathi, V; Praveen, K; Ravindran, P N; Peter, K V

    2016-01-01

    Ginger is a rhizomatous plant that belongs to the family Zingiberaceae. It is a herbaceous perennial but cultivated as annual, with crop duration of 7-10 months. Ginger is native to India and Tropical South Asia. The tuberous rhizomes or underground stems of ginger are used as condiment, an aromatic stimulant, and food preservative as well as in traditional medicine. Ginger is propagated vegetatively with rhizome bits as seed material. Cultivation of ginger is plagued by rhizome rot diseases, most of which are mainly spread through infected seed rhizomes. Micropropagation will help in production of disease-free planting material. Sexual reproduction is absent in ginger, making recombinant breeding very impossible. In vitro technology can thus become the preferred choice as it can be utilized for multiplication, conservation of genetic resources, generating variability, gene transfer, molecular tagging, and their utility in crop improvement of these crops.

  14. Agrobacterium-meditated Genetic Transformation of an Upland Cotton (Gossypium hirsutum cv Coker 310) Using a Novel Bt Gene Cry 2Ac

    Institute of Scientific and Technical Information of China (English)

    THIRUVENGADAM V; RASHMI J A; UDYASURIAN V; BALASUBRAMANIAN P; RAVEENDRAN T S

    2008-01-01

    @@ The development of transgenic cotton varieties resistant to bollworms has been a major success of applying plant genetic engineering technology to agriculture,evidenced by phenomenal increase in the cultivable area under (B.thuringiensis) Bt cotton in recent years worldwide.Of late,there are reports of insects developing resistance against the most commonly used Bt toxin CrylAc.Hence,there is an urgent need to broaden the source of resistance by employing new genes in order to reduce the chances of insects developing Bt resistance.Keeping this objective in view,cotton (Gossypium hirsuturn cv Coker 310) plants expressing a novel insecticidal crystal protein Cry2Ac were developed in the present study.

  15. Paper Genetic Engineering.

    Science.gov (United States)

    MacClintic, Scott D.; Nelson, Genevieve M.

    Bacterial transformation is a commonly used technique in genetic engineering that involves transferring a gene of interest into a bacterial host so that the bacteria can be used to produce large quantities of the gene product. Although several kits are available for performing bacterial transformation in the classroom, students do not always…

  16. Transformação genética: estratégias e aplicações para o melhoramento genético de espécies florestais Genetic transformation: strategies for forest species breeding

    Directory of Open Access Journals (Sweden)

    Laudete Maria Sartoretto

    2008-06-01

    Full Text Available A transformação genética, que consiste na introdução controlada de um gene no genoma de uma célula receptora e em sua posterior expressão, assume adicional significância, pois abre novas perspectivas ao melhoramento genético de espécies florestais, disponibilizando novos genes com características desejáveis para serem incorporados em menor espaço de tempo. Através do uso de estratégias com Agrobacterium e biobalística, já foram obtidas plantas transgênicas com maior produção de biomassa, melhor qualidade de madeira, maior resistência a determinados insetos e com tolerância a herbicidas, entre outras características de interesse. Tais exemplos demonstram a relevância dessas ferramentas para o setor florestal. Esta revisão apresenta, de forma resumida, a importância do estabelecimento de um eficiente sistema de regeneração in vitro, as principais estratégias usadas na transformação genética de espécies florestais e algumas das características agroflorestais que já foram incorporadas nessas espécies.The genetic transformation which consists in a controlled introduction of a gene in the genome of a receive cell and its subsequent expression, assumes additional significance, giving new perspectives to genetic improvement of the forest species, extending and available new genes with desirable characteristics, to be incorporated in a smaller period of time. Through the strategies Agrobacterium and bioballistic, transgenic plants had already been obtained showing higher biomass production, better wood quality, resistance to some insects, tolerance to herbicides, among other desirable characteristics, showing the relevance of these tools to the forest industry. This revision points out in a summarized way, the importance of establishing an efficient in vitro regeneration system, the strategies used in the genetic transformations of forestry species and some agroforestry characteristics that were previously incorporated

  17. Transformation of Anaplasma phagocytophilum

    Directory of Open Access Journals (Sweden)

    Barbet Anthony F

    2006-10-01

    Full Text Available Abstract Background Tick-borne pathogens cause emerging zoonoses, and include fastidious organisms such as Anaplasma phagocytophilum. Because of their obligate intracellular nature, methods for mutagenesis and transformation have not been available. Results To facilitate genetic manipulation, we transformed A. phagocytophilum (Ap to express a green fluorescent protein (GFP with the Himar1 transposase system and selection with the clinically irrelevant antibiotic spectinomycin. Conclusion These transformed bacteria (GFP/Ap grow at normal rates and are brightly fluorescent in human, monkey, and tick cell culture. Molecular characterization of the GFP/Ap genomic DNA confirmed transposition and the flanking genomic insertion locations were sequenced. Three mice inoculated with GFP/Ap by intraperitoneal injection became infected as demonstrated by the appearance of morulae in a peripheral blood neutrophil and re-isolation of the bacteria in culture.

  18. Ex vivo assays to study self-renewal, long-term expansion, and leukemic transformation of genetically modified human hematopoietic and patient-derived leukemic stem cells.

    Science.gov (United States)

    Sontakke, Pallavi; Carretta, Marco; Capala, Marta; Schepers, Hein; Schuringa, Jan Jacob

    2014-01-01

    With the emergence of the concept of the leukemic stem cell (LSC), assays to study them remain pivotal in understanding (leukemic) stem cell biology. Although the in vivo NOD-SCID or NSG xenotransplantation model is currently still the favored assay of choice in most cases, this system has some limitations as well such as its cost-effectiveness, duration, and lack of engraftability of cells from some acute myeloid leukemia (AML) patients. Here, we describe in vitro assays in which long-term expansion and self-renewal of LSCs isolated from AML patients can be evaluated. We have optimized lentiviral transduction procedures in order to stably express genes of interest or stably downmodulate genes using RNAi in primary AML cells, and these approaches are described in detail here. Also, we describe bone marrow stromal coculture systems in which cobblestone area-forming cell activity, self-renewal, long-term expansion, and in vitro myeloid or lymphoid transformation can be evaluated in human CD34(+) cells of fetal or adult origin that are engineered to express oncogenes. Together, these tools should allow a further molecular elucidation of derailed signal transduction in LSCs.

  19. An unusual abscisic acid and gibberellic acid synergism increases somatic embryogenesis, facilitates its genetic analysis and improves transformation in Medicago truncatula.

    Directory of Open Access Journals (Sweden)

    Kim E Nolan

    Full Text Available Somatic embryogenesis (SE can be readily induced in leaf explants of the Jemalong 2HA genotype of the model legume Medicago truncatula by auxin and cytokinin, but rarely in wild-type Jemalong. Gibberellic acid (GA, a hormone not included in the medium, appears to act in Arabidopsis as a repressor of the embryonic state such that low ABA (abscisic acid: GA ratios will inhibit SE. It was important to evaluate the GA effect in M. truncatula in order to formulate generic SE mechanisms, given the Arabidopsis information. It was surprising to find that low ABA:GA ratios in M. truncatula acted synergistically to stimulate SE. The unusual synergism between GA and ABA in inducing SE has utility in improving SE for regeneration and transformation in M. truncatula. Expression of genes previously shown to be important in M. truncatula SE was not increased. In investigating genes previously studied in GA investigations of Arabidopsis SE, there was increased expression of GA2ox and decreased expression of PICKLE, a negative regulator of SE in Arabidopsis. We suggest that in M. truncatula there are different ABA:GA ratios required for down-regulating the PICKLE gene, a repressor of the embryonic state. In M. truncatula it is a low ABA:GA ratio while in Arabidopsis it is a high ABA:GA ratio. In different species the expression of key genes is probably related to differences in how the hormone networks optimise their expression.

  20. 抗草甘膦基因的玉米转化及后代遗传分析%Transformation and Genetic Analysis of Maize with the Glyphosate-resistant Gene

    Institute of Scientific and Technical Information of China (English)

    王秀红; 史向远; 白建荣; 孙毅; 刘惠民

    2011-01-01

    利用农杆菌介导法将一个新型抗草甘膦基因导入玉米优良再生材料Hi-II中,分析基因在T0转化及后代的遗传情况,结果表明,在15株T0转基因植株中有10株,经PCR初步检测呈阳性;T1转基因植株用2%草甘膦进行抗筛选性后,4个穗行符合3:1的遗传分离比,3个穗行符合1:1的遗传分离比.对35个T2转基因植株以株系为单位混收进行PCR检测,结果表明,有32个T2株系表现阳性.对籽粒色泽纯度高且呈阳性的株系依次进行田间草甘膦涂抹、PCR和RT-PCR检测,结果显示,田间草甘膦筛选效果明显,植株全部存活,选10株PCR呈阳性的植株进行RT-PCR检测,9株出现1.2 kb的目的条带,说明抗草甘膦基因能够遗传至T2且能有效进行转录和翻译.%A new glyphosate-resistant gene was introduced into Hi-Ⅱ by Agrobacterium-mediated transformation,the To transformation and progeny genetics were analyzed. Ten plantlets were positive by PCR analysis from fifteen T0 transformed plants. After being painted with 2% glyphosate for 14 days,four spike lines accorded with 3:1 Mendelian segregation ratio and three spike lines accorded with 1:1 segregation ratio in T1. 32 spikes lines were positive by PCR from 35 T2 transformed plants. The spikes lines with high purity grain were selected to further glyphosate painting,PCR and RT-PCR analysis and all plants survived after glyphosate painting and 1.2 kb bands were obtained in nine from ten positive plants. The results showed that glyphosate-resistant gene was able to inherit to the T2 progeny,and transcribed and translated effectively.

  1. Eggplant (Solanum melongena L.: tissue culture, genetic transformation and use as an alternative model plant Berinjela (Solanum melongena L.: cultura de tecidos, transformação genética e uso como planta modelo

    Directory of Open Access Journals (Sweden)

    Claudia Magioli

    2005-03-01

    Full Text Available Eggplant is an agronomically important non-tuberous solanaceous crop grown primarily for its large oval fruit. In popular medicine, eggplant is indicated for the treatment of several diseases, including diabetes, arthritis, asthma and bronchitis. Eggplant is susceptible to a number of diseases and pests capable of causing serious crop losses. This problem has been addressed by hybridizing eggplant with wild resistant Solanum species, which present a wide genetic diversity and are source of useful agronomic traits. The application of in vitro methodologies to eggplant has resulted in considerable success. Eggplant tissues present a high morphogenetic potential that is useful for developmental studies as well as for establishing biotechnological approaches to produce improved varieties, such as embryo rescue, in vitro selection, somatic hybridization and genetic transformation. Taken together, these characteristics also make eggplant a complete model for studies on different areas of plant science, including control of gene expression and assessment of genetic stability of somaclones derived from different morphogenetic processes. In the present study, important factors that affect the efficiency of in vitro regeneration through organogenesis and embryogenesis as well as genetic transformation are analyzed. The potential of this species as a model plant for studying various aspects of plant genetics and physiology is also discussed.A berinjela é uma espécie solanácea não tuberosa de importância agronômica, cultivada principalmente por seus frutos. Na medicina popular, a berinjela é indicada para o tratamento de várias doenças, incluindo diabetes, artrite, asma e bronquite. A berinjela é suscetível a várias doenças e pragas que causam perdas econômicas significativas. Esse problema tem sido abordado com técnicas convencionais de melhoramento, utilizando espécies silvestres resistentes de Solanum, que possuem uma grande diversidade

  2. PLANT TRANSFORMATION: ADVANCES AND PERSPECTIVES

    Directory of Open Access Journals (Sweden)

    Alves Adriana Cristina

    1999-01-01

    Full Text Available Genetic transformation is a powerful tool for plant breeding and genetical, physiological or biochemical research, consequently it is an extremely dynamic field. Transgenic plants are commonly used to complete or substitute mutants in basic research, helping the studies of complex biological situations such as pathogenesis process, genome organization, light reception and signal transduction. In this review, recent approaches for foreign gene introduction (e.g. Agrobiolistics, whole tissue electroporation, in planta Agrobacterium transformation, screening (reporter gene possibilities and performance and transformant selection (ipt selective marker are discussed. Transgene expression and mechanisms underlying (transgene inactivation are presented. Practical applications of genetically modified plants, field tests and commercial transgenic crops worldwide and in Brazil are listed, as well as the main traits and species modified. Potential uses of transgenic plants for animal compound production, biological remediation and synthetic polymer assembly are also shown.

  3. Molecular genetic characterization of p53 mutated oropharyngeal squamous cell carcinoma cells transformed with human papillomavirus E6 and E7 oncogenes.

    Science.gov (United States)

    Oh, Ji-Eun; Kim, Jeong-Oh; Shin, Jung-Young; Zhang, Xiang-Hua; Won, Hye-Sung; Chun, Sang-Hoon; Jung, Chan-Kwon; Park, Won-Sang; Nam, Suk-Woo; Eun, Jung-Woo; Kang, Jin-Hyoung

    2013-08-01

    Patients with HPV-positive oropharyngeal cancer show better tumor response to radiation or chemotherapy than patients with HPV-negative cancer. HPV oncoprotein E6 binds and degrades a typically wild-type p53 protein product. However, HPV16 infection and p53 mutation infrequently coexist in a subset of HNSCCs. The purpose of this study was to investigate the mechanisms through which tumor biology and molecular genetic mechanisms change when two HPV-negative, p53-mutated oropharyngeal cell lines (YD8, non-disruptive p53 mutation; YD10B, disruptive p53 mutation) derived from patients with a history of heavy smoking are transfected with HPV E6 and E7 oncogenes in vitro. Transfection with HPV E6 and E7 oncogenes in YD8, reduced the abundance of proteins encoded by tumor suppressor genes, such as p-p53 and p-Rb. Cell proliferative activity was increased in the cells transfected with E6E7 compared to cells transfected with vector alone (P=0.09), whereas the invasiveness of E6E7-transfected cells was significantly reduced (P=0.02). cDNA microarray of the transfected cells with E6E7 showed significant changes in mRNA expression in several signaling pathways, including focal adhesion, JAK-STAT signaling pathway, cell cycle and p53 signaling pathway. Regarding the qPCR array for the p53 signaling pathway, the mRNA expression of STAT1 was remarkably upregulated by 6.47-fold (Pcell carcinoma cases with non-disruptive p53 mutations.

  4. 抗草甘膦基因mEPSPS转化油菜研究%Genetic transformation of rapeseed with the glyphosate-resistant gene mEPSPS

    Institute of Scientific and Technical Information of China (English)

    柳寒; 周永明

    2012-01-01

    Previously cloned mEPSPS gene [conferring glyphosate herbicide (Roundup) , resistance] was introduced into a Brassica napus line Jia 572 by Agrobacterium - mediated transformation method. Molecular analysis demonstrated that 4 independent transgenic plants were obtained, and the mEPSPS gene was inherited to the next generation. RT - PCR analysis indicated that the mEPSPS gene could be successfully transcribed in transgenic rapeseed plants although the expression levels varied among different lines. Herbicide resistance assay by Roundup spray showed that the transgenic plants survived under 100 - time diluted solution with commercial glyphosate -containing 41% herbicide (as 3 039mg/L glyphosate) , while the wild type died even under a 200 -time diluted solution.%mEPSPS基因是编码5-烯醇式丙酮酸莽草酸-3-磷酸合酶的抗草甘膦基因.本研究通过根癌农杆菌介导的遗传转化,将人工合成改造的草甘膦抗性基因mEPSPS导入甘蓝型油菜品系甲572,获得了4株转基因植株.分子检测证明,外源mEPSPS基因已整合到转基因油菜基因组中并能稳定遗传到下一代.各转基因植株中mEPSPS基因能正确表达,但不同转化株的基因表达量之间有差异.转mEPSPS油菜自交一代在稀释100倍的41%农达异丙胺盐制剂(含草甘膦3 039mg/L)喷洒条件下仍能正常生长,而不含转基因的对照植株在稀释200倍农达(含草甘膦1 519mg/L)之后全部死亡.

  5. 番茄NAM基因的克隆与遗传转化%Cloning and Genetic Transformation of NAM Gene in Tomato

    Institute of Scientific and Technical Information of China (English)

    杨春文; 唐宁; 李正国

    2012-01-01

    To understand the function of NAM gene in tomato, the full length cDNA NAM gene was amplified by RT-PCR from tomato cDNA, which contains a 1 053 bp open reading frame (ORF)encoding 351 amino acid residues, and contains NAC domain in its N terminus. Quantitative PCR analysis showed that NAM expressed highly in all tissues including root, stem, leaf, flower and fruit. To further investigate the function of tomato NAM gene, NAM overexpression vector pLP ?5S -NAM was constructed, and transformed into tomato plants by Agrobacterium-mediated method. The NAM overexpressing plants showed a dwarf phenotype, the size of these plants was smaller than those of wild-type plants, the plant growth was inhibited, and the leaves of transgenic tomato were highly lobed shaped. In conclusion, NAM affected the formation of shoot apical meristem(SAM)and leaf morphogenesis in tomato.%为了研究番茄NAM基因的功能,利用RT-PCR技术从番茄cDNA中分离了NAM(GenBank登录号:FJ435163.1)基因,该基因ORF全长1 053 bp,共编码351个氨基酸,N端含有NAC结构域.定量RT-PCR分析结果表明,该基因在番茄的根、茎、叶、花和果实中均有较强表达.为了进一步研究番茄NAM基因的功能,构建了NAM基因的超表达载体pLP100-35S-NAM,通过农杆菌介导法转化番茄,并获得抗性植株.经表型分析,转基因番茄植株矮小,生长受到抑制,叶片形态异常,表明NAM基因影响番茄顶端分生组织(SAM)的形成和叶片形态的发生.

  6. Visualizing Transformation

    DEFF Research Database (Denmark)

    Pedersen, Pia

    2012-01-01

    Transformation, defined as the step of extracting, arranging and simplifying data into visual form (M. Neurath, 1974), was developed in connection with ISOTYPE (International System Of TYpographic Picture Education) and might well be the most important legacy of Isotype to the field of graphic...... design. Recently transformation has attracted renewed interest because of the book ‘The Transformer’ written by Robin Kinross and Marie Neurath. My on-going research project, summarized in this paper, identifies and depicts the essential principles of data visualization underlying the process...... of transformation with reference to Marie Neurath’s sketches on the Bilston Project. The material has been collected at the Otto and Marie Neurath Collection housed at the University of Reading, UK. By using data visualization as a research method to look directly into the process of transformation the project...

  7. Transformational leadership.

    Science.gov (United States)

    Luzinski, Craig

    2011-12-01

    This month, the director of the Magnet Recognition Program® takes an in-depth look at the Magnet® model component transformational leadership. The author examines the expectations for Magnet organizations around this component. What are the qualities that make a nursing leader truly transformational, and what is the best approach to successfully lead a healthcare organization through today's volatile healthcare environment?

  8. Landskabets transformation

    DEFF Research Database (Denmark)

    Munck Petersen, Rikke

    2005-01-01

    Seminaroplæg fra forskere. Faglige seminarer på KA, forår 2005. Belyser transformation af det danske landskab fysisk som holdningsmæssigt, samt hvordan phd-arbejdets egen proces håndterer den.......Seminaroplæg fra forskere. Faglige seminarer på KA, forår 2005. Belyser transformation af det danske landskab fysisk som holdningsmæssigt, samt hvordan phd-arbejdets egen proces håndterer den....

  9. Establishment and Optimization of Genetic Transformation System of Lemon[Citrus limon (L.)Burm.f.]%柠檬遗传转化体系的建立及优化

    Institute of Scientific and Technical Information of China (English)

    仝铸; 何利刚; 吴黎明; 王志静; 蒋迎春; 孙中海

    2014-01-01

    以pCAMBIA1391Z为载体,对农杆菌EHA105介导的柠檬[Citrus limon (L.)Burm.f.]遗传转化体系进行了优化。结果表明,培养基中植物激素的浓度、农杆菌浸染浓度以及浸染后共培养的时间对柠檬的再生和遗传转化效率均有明显影响。1.0 mg/L 6-BA和0.1 mg/L的NAA有利于柠檬上胚轴不定芽的再生,并且再生芽的质量最佳。农杆菌的浸染浓度以OD600 nm=0.6为最佳,浸染后共培养2 d,在含5.0 mg/L潮霉素的筛选培养基上筛选,有利于柠檬不定芽再生率的提高。%Using pCAMBIA1391Z as vector, the genetic transformation system of lemon [Citrus limon (L.) Burm.f.]with A-grobacterium-mediated by EHA105 was optimized. The results showed that concentration of plant hormones, inoculum density and co-cultivation time had significant effects on lemon regeneration and transformation. It was most conducive to the regener-ation of adventitious buds lemon epicotyl by adding 1.0 mg/L 6-BA and 0.1 mg/L NAA in MT medium. The best inoculum density and co-cultivation time were 0.6 (OD600 nm) and two days. For the adventitious bud regeneration of lemon, the most benifical screening concentration of hygromycin was 5.0 mg/L.

  10. 基因组编辑技术及其在昆虫遗传转化中的应用%Genome editing techniques and their application in insect genetic transformation

    Institute of Scientific and Technical Information of China (English)

    吕志创; 严盈; 张桂芬; 武强; 刘万学; 万方浩

    2015-01-01

    The technologies of zinc finger nucleases ( ZFNs ) , transcription activator-like effector nucleases ( TALENs ) and CRISPR/Cas ( clustered regularly interspaced short palindromic repeats, CRISPR;CRISPR associated protein, Cas) are three ma-jor genome editing techniques that have being developed in recent years. The basic principle is to make DNA double-strand break ( DSB) damaged in the genome specific sites and to activate the body′s own DNA damage repair mechanism, then causing all sorts of variation in the process. The genome editing techniques have important research prospect in the research of gene function and gene repair. They are also expected to become the main strategy of insect genetic transformation since they have many merits. For exam-ple, CRISPR/Cas can be used to edit multiple specific loci in the genome. In this paper, the basic principles of the ZFNs, activa-tion of transcription factor type nucleic acid enzymes and CRISPR/Cas technologies and their applications in insects are briefly intro-duced. The information provides some references for the use of genome editing techniques in insect genetic transformation.%锌指核酸酶、类转录激活因子式核酸酶和CRISPR/Cas技术是近几年发展起来的3种主要基因组编辑技术,其原理都是通过在生物基因组特定位点制造DNA双链断裂损伤,从而激活机体自身的DNA 损伤修复机制,在此过程中引发各种变异。基因组编辑技术已在研究基因功能和基因修复中成功应用,基于基因组编辑技术的诸多优点,如CRISPR/Cas技术能对基因组中多个特定位点进行编辑,其有望成为昆虫遗传转化的主要策略。本文就锌指核酸酶、类转录激活因子式核酸酶和CRISPR/Cas技术的基本原理及其在昆虫中的应用做一简介,为今后利用基因组编辑技术进行昆虫遗传转化提供些许参考。

  11. A Better Method for SAR Image and Multi-Spectral Image Fusion Based on Nonsubsampled Contourlet Transform (NSCT) and Genetic Algorithm%基于NSCT和遗传算法的SAR图像和多光谱图像融合

    Institute of Scientific and Technical Information of China (English)

    时丕丽; 郭雷; 李晖晖; 杨宁; 陈智慧

    2012-01-01

    SAR image or multispectral image has big difference in imaging mechanism and spectral characteristics. Sections 1 and 2 of the full paper explain our image fusion method mentioned in the title, which we believe is new and better than previous ones. Their cpre consists of : (1) by using the multi-scale, multi-direction ad spare decomposition capability of the NSCT, we transform the SAR and multispectral source images into NSCT domain; (2) we fuse the low frequencey coefficients by maximizing the regional entropy; then we calculate the values of the high-frequence subband regional correlation coefficients, divide them into different ranges according to the threshold values selected by genetic algorithm and fuse the correlation coefficients in different ranges, ( 3) we take the inverse NSCT transform, thus obtatining the fused images. Section 3 simulates our image fusion method; the simulation results , given in Fig. 4 and Table 1, and their analysis show preliminarily that our image fusion method performs indeed much better than the exsiting regional-based/pixel-based Contourlet/NSCT.%合成孔径雷达(SAR)图像与多光谱图像成像机理和光谱特性差异较大,一般的融合方法很难取得满意的融合结果.文章提出了一种基于Nonsuosampled Contourlet transform(NSCT)和遗传算法的融合算法,首先将经过预处理后的图像进行NSCT分解,低频系数采取区域信息熵最大的准则融合;高频子带计算区域相关性,对相关性在不同阈值范围内的系数进行融合,阈值的选取采用遗传算法进行搜索;最后对融合系数进行NSCT逆变换,得到融合结果.仿真结果表明该算法显著优于基于像素点和基于区域的融合方法.

  12. Genetic parameters of cheese yield and curd nutrient recovery or whey loss traits predicted using Fourier-transform infrared spectroscopy of samples collected during milk recording on Holstein, Brown Swiss, and Simmental dairy cows.

    Science.gov (United States)

    Cecchinato, A; Albera, A; Cipolat-Gotet, C; Ferragina, A; Bittante, G

    2015-07-01

    Cheese yield is the most important technological parameter in the dairy industry in many countries. The aim of this study was to infer (co)variance components for cheese yields (CY) and nutrient recoveries in curd (REC) predicted using Fourier-transform infrared (FTIR) spectroscopy of samples collected during milk recording on Holstein, Brown Swiss, and Simmental dairy cows. A total of 311,354 FTIR spectra representing the test-day records of 29,208 dairy cows (Holstein, Brown Swiss, and Simmental) from 654 herds, collected over a 3-yr period, were available for the study. The traits of interest for each cow consisted of 3 cheese yield traits (%CY: fresh curd, curd total solids, and curd water as a percent of the weight of the processed milk), 4 curd nutrient recovery traits (REC: fat, protein, total solids, and the energy of the curd as a percent of the same nutrient in the processed milk), and 3 daily cheese production traits (daily fresh curd, total solids, and the water of the curd per cow). Calibration equations (freely available upon request to the corresponding author) were used to predict individual test-day observations for these traits. The (co)variance components were estimated for the CY, REC, milk production, and milk composition traits via a set of 4-trait analyses within each breed. All analyses were performed using REML and linear animal models. The heritabilities of the %CY were always higher for Holstein and Brown Swiss cows (0.22 to 0.33) compared with Simmental cows (0.14 to 0.18). In general, the fresh cheese yield (%CYCURD) showed genetic variation and heritability estimates that were slightly higher than those of its components, %CYSOLIDS and %CYWATER. The parameter RECPROTEIN was the most heritable trait in all the 3 breeds, with values ranging from 0.32 to 0.41. Our estimation of the genetic relationships of the CY and REC with milk production and composition revealed that the current selection strategies used in dairy cattle are expected

  13. Transformação genética de laranja 'Valência' com o gene cecropin MB39 Genetic transformation of 'Valencia' sweet orange with the cecropin MB39 gene

    Directory of Open Access Journals (Sweden)

    Luis Gustavo de Paoli

    2007-11-01

    Full Text Available O objetivo deste trabalho foi obter plantas transgênicas de laranja 'Valência' com o gene cecropin MB39 controlado pelo promotor do gene da fenilalanina-amônia-liase de citros, visando a expressão gênica específica nos vasos do xilema. A transformação genética foi realizada via Agrobacterium tumefaciens por meio do co-cultivo de segmentos de epicótilo. Onze plantas transgênicas foram identificadas por PCR, pela amplificação do fragmento esperado de 189 pb, as quais foram aclimatizadas em casa de vegetação. A integração do transgene foi confirmada em três plantas pela análise de transferência de Southern.The objective of this work was to produce 'Valencia' sweet orange transgenic plants with the cecropin MB39 gene controlled by a phenylalanine ammonia-lyase gene promoter from citrus in order to direct gene expression in xylem vessels. The genetic transformation was mediated by Agrobacterium tumefaciens with the co-culture of epicotyl segments. Eleven transgenic plants were selected by PCR with the amplification of a 189 bp fragment, which were acclimatized to greenhouse. The integration of the transgene was confirmed in three plants by Southern blot analysis.

  14. Foundations of genetic algorithms 1991

    CERN Document Server

    1991-01-01

    Foundations of Genetic Algorithms 1991 (FOGA 1) discusses the theoretical foundations of genetic algorithms (GA) and classifier systems.This book compiles research papers on selection and convergence, coding and representation, problem hardness, deception, classifier system design, variation and recombination, parallelization, and population divergence. Other topics include the non-uniform Walsh-schema transform; spurious correlations and premature convergence in genetic algorithms; and variable default hierarchy separation in a classifier system. The grammar-based genetic algorithm; condition

  15. Identity transformation

    DEFF Research Database (Denmark)

    Neergaard, Helle; Robinson, Sarah; Jones, Sally

    This paper develops the concept of ‘pedagogical nudging’ and examines four interventions in an entrepreneurship classroom and the potential it has for student identity transformation. Pedagogical nudging is positioned as a tool, which in the hands of a reflective, professional, with an understand......This paper develops the concept of ‘pedagogical nudging’ and examines four interventions in an entrepreneurship classroom and the potential it has for student identity transformation. Pedagogical nudging is positioned as a tool, which in the hands of a reflective, professional......, as well as the resources they have when they come to the classroom. It also incorporates perspectives from (ii) transformational learning and explores the concept of (iii) nudging from a pedagogical viewpoint, proposing it as an important tool in entrepreneurship education. The study incorporates......) assists students in straddling the divide between identities, the emotions and tensions this elicits, and (iv) transform student understanding. We extend nudging theory into a new territory. Pedagogical nudging techniques may be able to unlock doors and bring our students beyond the unacknowledged...

  16. Sustainable transformation

    DEFF Research Database (Denmark)

    Andersen, Nicolai Bo

    , that it can be adapted to changing functional needs, and that it has an architectural and cultural value. A specific proposal for a transformation that enhances the architectural qualities and building heritage values of an existing building forms the empirical material, which is discussed using different...

  17. ADE Transform

    CERN Document Server

    Donagi, Ron

    2015-01-01

    There is a beautiful correspondence between configurations of lines on a rational surface and tautological bundles over that surface. We extend this correspondence to families, by means of a generalized Fourier-Mukai transform that relates spectral data to bundles over a rational surface fibration.

  18. Transformer core

    NARCIS (Netherlands)

    Mehendale, A.; Hagedoorn, Wouter; Lötters, Joost Conrad

    2010-01-01

    A transformer core includes a stack of a plurality of planar core plates of a magnetically permeable material, which plates each consist of a first and a second sub-part that together enclose at least one opening. The sub-parts can be fitted together via contact faces that are located on either side

  19. Transformer core

    NARCIS (Netherlands)

    Mehendale, A.; Hagedoorn, Wouter; Lötters, Joost Conrad

    2008-01-01

    A transformer core includes a stack of a plurality of planar core plates of a magnetically permeable material, which plates each consist of a first and a second sub-part that together enclose at least one opening. The sub-parts can be fitted together via contact faces that are located on either side

  20. Transformational change

    NARCIS (Netherlands)

    Termeer, Katrien; Dewulf, Art; Biesbroek, Robbert

    2016-01-01

    Although transformational change is a rather new topic in climate change adaptation literature, it has been studied in organisational theory for over 30 years. This paper argues that governance scholars can learn much from organisation theory, more specifically regarding the conceptualisation of

  1. Comparison between genetic parameters of cheese yield and nutrient recovery or whey loss traits measured from individual model cheese-making methods or predicted from unprocessed bovine milk samples using Fourier-transform infrared spectroscopy.

    Science.gov (United States)

    Bittante, G; Ferragina, A; Cipolat-Gotet, C; Cecchinato, A

    2014-10-01

    Cheese yield is an important technological trait in the dairy industry. The aim of this study was to infer the genetic parameters of some cheese yield-related traits predicted using Fourier-transform infrared (FTIR) spectral analysis and compare the results with those obtained using an individual model cheese-producing procedure. A total of 1,264 model cheeses were produced using 1,500-mL milk samples collected from individual Brown Swiss cows, and individual measurements were taken for 10 traits: 3 cheese yield traits (fresh curd, curd total solids, and curd water as a percent of the weight of the processed milk), 4 milk nutrient recovery traits (fat, protein, total solids, and energy of the curd as a percent of the same nutrient in the processed milk), and 3 daily cheese production traits per cow (fresh curd, total solids, and water weight of the curd). Each unprocessed milk sample was analyzed using a MilkoScan FT6000 (Foss, Hillerød, Denmark) over the spectral range, from 5,000 to 900 wavenumber × cm(-1). The FTIR spectrum-based prediction models for the previously mentioned traits were developed using modified partial least-square regression. Cross-validation of the whole data set yielded coefficients of determination between the predicted and measured values in cross-validation of 0.65 to 0.95 for all traits, except for the recovery of fat (0.41). A 3-fold external validation was also used, in which the available data were partitioned into 2 subsets: a training set (one-third of the herds) and a testing set (two-thirds). The training set was used to develop calibration equations, whereas the testing subsets were used for external validation of the calibration equations and to estimate the heritabilities and genetic correlations of the measured and FTIR-predicted phenotypes. The coefficients of determination between the predicted and measured values in cross-validation results obtained from the training sets were very similar to those obtained from the whole

  2. Genetic Discrimination

    Science.gov (United States)

    ... in Genetics Archive Regulation of Genetic Tests Genetic Discrimination Overview Many Americans fear that participating in research ... I) and employment (Title II). Read more Genetic Discrimination and Other Laws Genetic Discrimination and Other Laws ...

  3. Evaluation of two cotton varieties CRSP1 and CRSP2 for genetic transformation efficiency, expression of transgenes Cry1Ac + Cry2A, GT gene and insect mortality

    Directory of Open Access Journals (Sweden)

    Arfan Ali

    2016-03-01

    Full Text Available Expression of the transgene with a desirable character in crop plant is the ultimate goal of transgenic research. Transformation of two Bt genes namely Cry1Ac and Cry2A cloned as separate cassette under 35S promoter in pKHG4 plant expression vector was done by using shoot apex cut method of Agrobacterium. Molecular confirmation of putative transgenic cotton plants for Cry1Ac, Cry2A and GT gene was done through PCR and ELISA. Transformation efficiency of CRSP-1 and CRSP-2 was calculated to be 1.2 and 0.8% for Cry1Ac while 0.9 and 0.6% for Cry2A and 1.5 and 0.7% for GTG respectively. CRSP-1 was found to adopt natural environment (acclimatized earlier than CRSP-2 when exposed to sunlight for one month. Expression of Cry1Ac, Cry2A and GTG was found to be 1.2, 1 and 1.3 ng/μl respectively for CRSP-1 as compared to CRSP-2 where expression was recorded to be 0.9, 0.5 and 0.9 ng/μl respectively. FISH analysis of the transgenic CRSP-1 and CRSP-2 demonstrated the presence of one and two copy numbers respectively. Similarly, the response of CRSP-1 against Glyphosate @1900 ml/acre was far better with almost negligible necrotic spot and efficient growth after spray as compared to CRSP-2 where some plants were found to have necrosis and negative control where the complete decay of plant was observed after seven days of spray assay. Similarly, almost 100% mortality of 2nd instar larvae of Heliothis armigera was recorded after three days in CRSP-1 as compared CRSP-2 where insect mortality was found to be less than 90%. Quantitatively speaking non transgenic plants were found with 23–90% leaf damage by insect, while CRSP-1 was with less than 5% and CRSP-2 with 17%. Taken together CRSP1 was found to have better insect control and weedicide resistance along with its natural ability of genetic modification and can be employed by the valuable farmers for better insect control and simultaneously for better production.

  4. Plant Regeneration and Genetic Transformation in Eggplant ...

    African Journals Online (AJOL)

    Dr Harmander Gill

    2014-02-05

    Feb 5, 2014 ... 2School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana, 141004 India. Accepted 5 ... media affect its regeneration potential (Kantharajah and .... Rogor and Kitazin in relation to their concentrations also.

  5. [Transformation of genetically engineered oxytocinoyllysine into oxytocin].

    Science.gov (United States)

    Mal'tsev, K V; Shvets, S V; Luk'ianova, N A; Grigor'ev, A V; Miroshnikov, A I

    1996-04-01

    Various methods for obtaining oxytocin from its recombinant precursor oxytocinoyllysine were studied. For splitting off the C-terminal lysine residue in oxytocinoyllysine, the carboxypeptidase B and an analogous carboxypeptidase from king crab hepatopancreas were used. Ammonolysis of oxytocinic acid methyl ester proved to be the most efficient method for the last stage of the oxytocin preparation. Reversed-phase HPLC was used for the product analysis at each stage of the recombinant oxytocinoyllysine conversion into oxytocin.

  6. Discrete transforms

    CERN Document Server

    Firth, Jean M

    1992-01-01

    The analysis of signals and systems using transform methods is a very important aspect of the examination of processes and problems in an increasingly wide range of applications. Whereas the initial impetus in the development of methods appropriate for handling discrete sets of data occurred mainly in an electrical engineering context (for example in the design of digital filters), the same techniques are in use in such disciplines as cardiology, optics, speech analysis and management, as well as in other branches of science and engineering. This text is aimed at a readership whose mathematical background includes some acquaintance with complex numbers, linear differen­ tial equations, matrix algebra, and series. Specifically, a familiarity with Fourier series (in trigonometric and exponential forms) is assumed, and an exposure to the concept of a continuous integral transform is desirable. Such a background can be expected, for example, on completion of the first year of a science or engineering degree cour...

  7. XML Transformations

    Directory of Open Access Journals (Sweden)

    Felician ALECU

    2012-04-01

    Full Text Available XSLT style sheets are designed to transform the XML documents into something else. The two most popular parsers of the moment are the Document Object Model (DOM and the Simple API for XML (SAX. DOM is an official recommendation of the W3C (available at http://www.w3.org/TR/REC-DOM-Level-1, while SAX is a de facto standard. A good parser should be fast, space efficient, rich in functionality and easy to use.

  8. Transformative Agency

    DEFF Research Database (Denmark)

    Majgaard, Klaus

    The purpose of this paper is to enhance the conceptual understanding of the mediatory relationship between paradoxes on an organizational and an individual level. It presents a concept of agency that comprises and mediates between a structural and individual pole. The constitution of this agency ...... is achieved through narrative activity that oscillates between the poles and transforms paradoxes through the configuration of plots and metaphors. Empirical cases are introduced in order to illustrate the implications of this understanding....

  9. RF transformer

    Science.gov (United States)

    Smith, James L.; Helenberg, Harold W.; Kilsdonk, Dennis J.

    1979-01-01

    There is provided an improved RF transformer having a single-turn secondary of cylindrical shape and a coiled encapsulated primary contained within the secondary. The coil is tapered so that the narrowest separation between the primary and the secondary is at one end of the coil. The encapsulated primary is removable from the secondary so that a variety of different capacity primaries can be utilized with one secondary.

  10. Subspaces of FMmlet transform

    Institute of Scientific and Technical Information of China (English)

    邹红星; 戴琼海; 赵克; 陈桂明; 李衍达

    2002-01-01

    The subspaces of FMmlet transform are investigated.It is shown that some of the existing transforms like the Fourier transform,short-time Fourier transform,Gabor transform,wavelet transform,chirplet transform,the mean of signal,and the FM-1let transform,and the butterfly subspace are all special cases of FMmlet transform.Therefore the FMmlet transform is more flexible for delineating both the linear and nonlinear time-varying structures of a signal.

  11. Relations of FMmlet Transform to Some Integral Transforms

    Institute of Scientific and Technical Information of China (English)

    ZOUHongxing; DAIQionghai; WANGDianjun; LIYanda

    2004-01-01

    In this paper, we explore the relationships between FMmlet transform and some of the existing integral transforms, namely, the chirplet transform, dispersion transform, wavelet transform, chirp-Fourier transform, Short-time fourier transform (STFT), Gabor transform, Fourier transform, cosine transform, sine transform,Hartley transform, Laplace transform, z-transform, Mellintransform, Hilbert transform, autocorrelation function,cross-correlation function, energy, and the mean value.It is shown that all of these transforms are subspaces of FMmlet transform with specific parameters.

  12. Hamlet's Transformation.

    Science.gov (United States)

    Usher, P. D.

    1997-12-01

    William Shakespeare's Hamlet has much evidence to suggest that the Bard was aware of the cosmological models of his time, specifically the geocentric bounded Ptolemaic and Tychonic models, and the infinite Diggesian. Moreover, Shakespeare describes how the Ptolemaic model is to be transformed to the Diggesian. Hamlet's "transformation" is the reason that Claudius, who personifies the Ptolemaic model, summons Rosencrantz and Guildenstern, who personify the Tychonic. Pantometria, written by Leonard Digges and his son Thomas in 1571, contains the first technical use of the word "transformation." At age thirty, Thomas Digges went on to propose his Perfit Description, as alluded to in Act Five where Hamlet's age is given as thirty. In Act Five as well, the words "bore" and "arms" refer to Thomas' vocation as muster-master and his scientific interest in ballistics. England's leading astronomer was also the father of the poet whose encomium introduced the First Folio of 1623. His oldest child Dudley became a member of the Virginia Company and facilitated the writing of The Tempest. Taken as a whole, such manifold connections to Thomas Digges support Hotson's contention that Shakespeare knew the Digges family. Rosencrantz and Guildenstern in Hamlet bear Danish names because they personify the Danish model, while the king's name is latinized like that of Claudius Ptolemaeus. The reason Shakespeare anglicized "Amleth" to "Hamlet" was because he saw a parallel between Book Three of Saxo Grammaticus and the eventual triumph of the Diggesian model. But Shakespeare eschewed Book Four, creating this particular ending from an infinity of other possibilities because it "suited his purpose," viz. to celebrate the concept of a boundless universe of stars like the Sun.

  13. Rotary Transformer

    Science.gov (United States)

    McLyman, Colonel Wm. T.

    1996-01-01

    None given. From first Par: Many spacecraft (S/C) and surface rovers require the transfer of signals and power across rotating interfaces. Science instruments, antennas and solar arrays are elements needing rotary power transfer for certain (S/C) configurations. Delivery of signal and power has mainly been done by using the simplest means, the slip ring approach. This approach, although simple, leaves debris generating noise over a period of time...The rotary transformer is a good alternative to slip rings for signal and power transfer.

  14. TRANSFORMER APPARATUS

    Science.gov (United States)

    Wolfgang, F.; Nicol, J.

    1962-11-01

    Transformer apparatus is designed for measuring the amount of a paramagnetic substance dissolved or suspended in a diamagnetic liquid. The apparatus consists of a cluster of tubes, some of which are closed and have sealed within the diamagnetic substance without any of the paramagnetic material. The remaining tubes are open to flow of the mix- ture. Primary and secondary conductors are wrapped around the tubes in such a way as to cancel noise components and also to produce a differential signal on the secondaries based upon variations of the content of the paramagnetic material. (AEC)

  15. Transformational leadership.

    Science.gov (United States)

    Marlow, D L

    1996-01-01

    In these uncertain times in the healthcare industry, administrators are asked to do more with less time and resources. Because of the extended roles they are playing in today's organizations, radiology administrators are looked upon as agents of change. What leadership skills do they need in this turbulent and uncertain healthcare environment? What are the trait's of tomorrow's leaders? The transformational leader is the one who will guide us through this changing healthcare environment. Several behavioral patterns emerge as important traits for tomorrow's leaders to have-individual consideration, intellectual stimulation and charisma. Tomorrow's leader must view each person as an individual, showing genuine concern and belief in each person's ability to perform. Transformational leaders stimulate others by encouraging them to be curious and try new ideas. The final characteristic, charisma, is the ability to inspire others. Luckily, leaders are made, not born: today's leaders can learn to be responsive, to draw out new ideas from employees, and to communicate self-esteem, energy and enthusiasm.

  16. New Genetics

    Science.gov (United States)

    ... Home > Science Education > The New Genetics The New Genetics Living Laboratories Classroom Poster Order a Free Copy ... Piece to a Century-Old Evolutionary Puzzle Computing Genetics Model Organisms RNA Interference The New Genetics is ...

  17. An optimized transformation protocol for Lipomyces starkeyi.

    Science.gov (United States)

    Calvey, Christopher H; Willis, Laura B; Jeffries, Thomas W

    2014-08-01

    We report the development of an efficient genetic transformation system for Lipomyces starkeyi based on a modified lithium acetate transformation protocol. L. starkeyi is a highly lipogenic yeast that grows on a wide range of substrates. The initial transformation rate for this species was extremely low, and required very high concentrations of DNA. A systematic approach for optimizing the protocol resulted in an increase in the transformation efficiency by four orders of magnitude. Important parameters included cell density, the duration of incubation and recovery periods, the heat shock temperature, and the concentration of lithium acetate and carrier DNA within the transformation mixture. We have achieved efficiencies in excess of 8,000 transformants/µg DNA, which now make it possible to screen libraries in the metabolic engineering of this yeast. Metabolic engineering based on this transformation system could improve lipogenesis and enable formation of higher value products.

  18. Cancer: Some genetic considerations

    African Journals Online (AJOL)

    Mohammad Saad Zaghloul Salem

    2014-09-22

    Sep 22, 2014 ... quite reasonable, as cancer is primarily a genetic alteration, lack of .... induced by malignant transformation impart to the cell new ... involved in this mysterious biological behavior of cells. ... the preservation of the evolutionary ability of the zygote ... by genomic regulatory mechanisms controlled by master.

  19. Medical Genetics Is Not Eugenics

    Science.gov (United States)

    Cowan, Ruth Schwartz

    2008-01-01

    The connection that critics make between medical genetics and eugenics is historically fallacious. Activists on the political right are as mistaken as activists on the political left: Genetic screening was not eugenics in the past, is not eugenics in the present, and, unless its technological systems become radically transformed, will not be…

  20. Genomic simulation for the study of data transformation efficiency in the genetic evaluation of cattle in the presence of heterogeneity=Simulação genômica para estudo da eficiência da transformação de dados para avaliação genética de bovinos na presença de heterogeneidade

    Directory of Open Access Journals (Sweden)

    Fabyano Fonseca e Silva

    2012-01-01

    Full Text Available Efficiency of data transformation strategies is evaluated to correct heterogeneity among cattle in the genetic evaluation of bulls, cows and progenies. Four data structures of bovine weaning weights were simulated: herds with and without heterogeneity for means and variances, and herds with and without genetic connectedness. In the genetic evaluations, data were used in the original scale and transformed (Logarithmic, Square root, standardization and ratio by phenotypic standard deviation. The evaluated data transformation strategies were not efficient in eliminating the negative effects of heterogeneity among cattle in the genetic evaluation of bulls, cows and progenies.O objetivo deste estudo foi avaliar a eficiência de estratégias de transformação de dados para correção da heterogeneidade entre rebanhos na avaliação genética de touros, vacas e progênies. Quatro estruturas de dados relativos a peso à desmama de bovinos foram simuladas: rebanhos com e sem heterogeneidade para médias e variâncias, e rebanhos com e sem conexidade genética. Nas avaliações genéticas, utilizaram-se dados na escala original e transformados (Logarítmica, Raiz quadrada, Padronização e Razão pelo desvio-padrão fenotípico. As estratégias de transformação de dados avaliadas não foram eficientes para eliminar os efeitos negativos da heterogeneidade entre rebanhos sobre a avaliação genética de touros, vacas e progênies.

  1. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation.

    Science.gov (United States)

    Romero, Diego; Pérez-García, Alejandro; Veening, Jan-Willem; de Vicente, Antonio; Kuipers, Oscar P

    2006-09-01

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool for molecular genetic analysis of interesting Bacillus strains, which are hard to transform by conventional methods.

  2. Fluorescence Assisted Selection of Transformants (FAST): Using flow cytometry to select fungal transformants

    NARCIS (Netherlands)

    Vlaardingerbroek, I.; Beerens, B.; Shahi, S.; Rep, M.

    2015-01-01

    The availability of drug resistance markers for fungal transformation is often a limiting factor in both fungal genetics research and industrial applications. We describe a new technique using flow cytometry to select fungal transformants using well-known fluorescent proteins as markers for transfor

  3. Agrobacterium-mediated transformation of barley (Hordeum vulgare L.).

    Science.gov (United States)

    Ismagul, Ainur; Mazonka, Iryna; Callegari, Corinne; Eliby, Serik

    2014-01-01

    Barley biotechnology requires efficient genetic engineering tools for producing transgenic plants necessary for conducting reverse genetics analyses in breeding and functional genomics research. Agrobacterium-mediated genetic transformation is an important technique for producing barley transgenics with simple low-copy number transgenes. This chapter reports a refined protocol for the systematic high-throughput transformation of the advanced Australian spring barley breeding line WI4330.

  4. [Agrobacterium-mediated transformation of Cymbidium sinensis].

    Science.gov (United States)

    Xie, Li; Wang, Fen; Zeng, Ruizhen; Guo, Herong; Zhou, Yuliang; Zhang, Zhisheng

    2015-04-01

    Genetic transformation is an effective method to improve breeding objective traits of orchids. However, there is little information about genetic transformation of Cymbidium sinensis. Rhizomes from shoot-tip culture of C. sinensis cv. 'Qijianbaimo' were used to establish a practical transformation protocol of C. sinensis. Pre-culture time, concentration and treating methods of acetosyringone, concentration of infection bacteria fluid (OD600), infection time, and co-culture time had significant effects on β-glucuronidase (GUS) transient expression rate of C. sinensis cv. 'Qijianbaimo' rhizome. The GUS transient expression rate of rhizome was the highest (11.67%) when rhizomes pre-cultured for 39 d were soaked in bacterium suspension (OD600 = 0.9) supplemented with 200 μmol/L acetosyringone for 35 min, followed by culturing on co-culture medium supplemented with 200 μmol/L acetosyringone for 7 d. Under this transformation conditions, 3 transgenic plantlets, confirmed by GUS histochemical assay and PCR, were obtained from 400 regenerated plantlets, and the genetic transformation rate was 0.75%. This proved that it was feasible to create new cultivars by the use of Agrobacterium-mediated genetic transformation in C. sinense.

  5. Genetic algorithms

    Science.gov (United States)

    Wang, Lui; Bayer, Steven E.

    1991-01-01

    Genetic algorithms are mathematical, highly parallel, adaptive search procedures (i.e., problem solving methods) based loosely on the processes of natural genetics and Darwinian survival of the fittest. Basic genetic algorithms concepts are introduced, genetic algorithm applications are introduced, and results are presented from a project to develop a software tool that will enable the widespread use of genetic algorithm technology.

  6. Genetic Mapping

    Science.gov (United States)

    ... Fact Sheets Fact Sheets En Español: Mapeo Genético Genetic Mapping What is genetic mapping? How do researchers create ... genetic map? What are genetic markers? What is genetic mapping? Among the main goals of the Human Genome ...

  7. Transforming giants.

    Science.gov (United States)

    Kanter, Rosabeth Moss

    2008-01-01

    Large corporations have long been seen as lumbering, inflexible, bureaucratic--and clueless about global developments. But recently some multinationals seem to be transforming themselves: They're engaging employees, moving quickly, and introducing innovations that show true connection with the world. Harvard Business School's Kanter ventured with a research team inside a dozen global giants--including IBM, Procter & Gamble, Omron, CEMEX, Cisco, and Banco Real--to discover what has been driving the change. After conducting more than 350 interviews on five continents, she and her colleagues came away with a strong sense that we are witnessing the dawn of a new model of corporate power: The coordination of actions and decisions on the front lines now appears to stem from widely shared values and a sturdy platform of common processes and technology, not from top-down decrees. In particular, the values that engage the passions of far-flung workforces stress openness, inclusion, and making the world a better place. Through this shift in what might be called their guidance systems, the companies have become as creative and nimble as much smaller ones, even while taking on social and environmental challenges of a scale that only large enterprises could attempt. IBM, for instance, has created a nonprofit partnership, World Community Grid, through which any organization or individual can donate unused computing power to research projects and see what is being done with the donation in real time. IBM has gained an inspiring showcase for its new technology, helped business partners connect with the company in a positive way, and offered individuals all over the globe the chance to contribute to something big.

  8. Antibiotic resistance of Nannochloropsis oculata and determination of selection markers for its genetic transformation%微拟球藻抗生素抗性分析和选择标记筛选

    Institute of Scientific and Technical Information of China (English)

    张琳; 马晓磊; 朱葆华; 于文功; 杨官品; 潘克厚

    2011-01-01

    为了筛选到微拟球藻Nannochloropsis oculata的筛选标记,研究了微拟球藻对新霉素(Neomycin)、链霉素(Streptomycin)、潮霉素(Hygromycin)、壮观霉素(Spectinomycin)和博来霉素(Zeocin)的敏感性.每种抗生素都设置液体培养和固体培养实验并且浓度梯度相同.新霉素和链霉素的浓度梯度为:0、100、200、400、600、800 μg/ml.潮霉素的浓度梯度为0、25、50、100、125、150μg/ml.壮观霉素浓度梯度为0、40、80、100、150、200 μg/ml.博来霉素浓度梯度则为0、0.1、0.2、0.4、0.5、1μg/ml.结果表明,微拟球藻对新霉素、链霉素、潮霉素和壮观霉素不敏感,而对博来霉素高度敏感.在固体培养时,0.5 μg/ml博来霉素即可完全抑制微拟球藻生长;在液体培养时,1 μg/ml博来霉素即可完全抑制其生长.博来霉素可作为微拟球藻遗传转化的选择抗生素.ble基因为其阳性筛选的选择标记基因.%To determine the selection marker of N. Oculata, the sensitivity to 5 antibiotics (Neomycin, Streptomycin, Hygromycin, Spectinomycin, and Zeocin) was determined in this study. For each antibiotic, liquid medium and solid medium were both adopted with the same concentration gradient. The concentration gradient of Neomycin and Streptomycin was 0, 100, 200, 400, 600, and 800μg/ml, while that of Hygromycin was 0, 25, 50, 100, 125, and 150μg/ ml, and that of Spectinomycin was 0, 40, 80, 100, 150, and 200μg/ml. The concentrations of Zeocin were much lower, which were 0, 0.1, 0. 2, 0. 4, 0. 5, and μg/ml. It was found that N. Oculata is not sensitive to Neomycin, Streptomycin, Hygromycin or Spectinomycin, but highly sensitive to Zeocin. Zeocin can completely inhibit the growth of N. Oculata when its con centration is 0. 5μg/ml in solid medium or 1μg/ml in liquid medium. Zeocin could serve as a se lection reagent in the genetic transformation of N. Oculata, and the ble gene could be the selec tion marker.

  9. Organogenesis and transient genetic transformation of the hybrid Eucalyptus grandis × Eucalyptus urophylla Organogênese e transformação genética transiente do híbrido Eucalyptus grandis × Eucalyptus urophylla

    Directory of Open Access Journals (Sweden)

    Giovana Bomfim de Alcantara

    2011-04-01

    Full Text Available The hybrid Eucalyptus grandis x Eucalyptus urophylla presents high levels of productivity and potential for use in paper, cellulose and fiber industries. The bud organogenesis from leaf explants of two clones of E. grandis × E. urophylla was studied in order to verify the effect of several factors: subculture duration on multiplication medium, type of explants, entire and half leaves: basal and apical portions, and duration of the culture on a regeneration medium. Differences in organogenic capacity of the two clones tested were observed. The explant most recommended for organogenesis is the basal section of the leaf collected from shoot clusters subcultured every 17 days. Moreover, the leaf explants must be transferred to a fresh bud induction medium every five days. This study also aimed at evaluating factors affecting the genetic transformation of leaf explants with the uidA gene, via co-cultivation with Agrobacterium tumefaciens, such as the pre-culture of the explants on a specific medium, the duration of their co-culture with the bacteria and the addition of acetosyringone to the culture media. The best conditions for the expression of the uidA gene were two days of pre-culture of the leaf tissues, three days of co-culture with the bacteria and the addition of acetosyringone in pre- and co-culture media.O híbrido Eucalyptus grandis x Eucalyptus urophylla apresenta alta produtividade e potencial para indústrias de papel, celulose e fibras. A organogênese de explantes foliares de dois clones de E. grandis × E. urophylla foi estudada para verificar fatores como tempo de repicagem das plântulas matrizes em meio de multiplicação; tipo de explantes, folhas inteiras e de meias-folhas (porções basais e apicais e dos dias que os explantes foliares permaneceram em meio de regeneração. Foram observadas diferenças na capacidade organogênica dos dois clones testados. A parte basal das folhas, coletadas de brotações repicadas a cada 17

  10. Genetic Counseling

    Science.gov (United States)

    Genetic counseling provides information and support to people who have, or may be at risk for, genetic disorders. A ... meets with you to discuss genetic risks. The counseling may be for yourself or a family member. ...

  11. Plastid transformation in sugar beet: Beta vulgaris.

    Science.gov (United States)

    De Marchis, Francesca; Bellucci, Michele

    2014-01-01

    Chloroplast biotechnology has assumed great importance in the past 20 years and, thanks to the numerous advantages as compared to conventional transgenic technologies, has been applied in an increasing number of plant species but still very much limited. Hence, it is of utmost importance to extend the range of species in which plastid transformation can be applied. Sugar beet (Beta vulgaris L.) is an important industrial crop of the temperate zone in which chloroplast DNA is not transmitted trough pollen. Transformation of the sugar beet genome is performed in several research laboratories; conversely sugar beet plastome genetic transformation is far away from being considered a routine technique. We describe here a method to obtain transplastomic sugar beet plants trough biolistic transformation. The availability of sugar beet transplastomic plants should avoid the risk of gene flow between these cultivated genetic modified sugar beet plants and the wild-type plants or relative wild species.

  12. Evolution and genetic diversity of Theileria.

    Science.gov (United States)

    Sivakumar, Thillaiampalam; Hayashida, Kyoko; Sugimoto, Chihiro; Yokoyama, Naoaki

    2014-10-01

    Theileria parasites infect a wide range of domestic and wild ruminants worldwide, causing diseases with varying degrees of severity. A broad classification, based on the parasite's ability to transform the leukocytes of host animals, divides Theileria into two groups, consisting of transforming and non-transforming species. The evolution of transforming Theileria has been accompanied by drastic changes in its genetic makeup, such as acquisition or expansion of gene families, which are thought to play critical roles in the transformation of host cells. Genetic variation among Theileria parasites is sometimes linked with host specificity and virulence in the parasites. Immunity against Theileria parasites primarily involves cell-mediated immune responses in the host. Immunodominance and major histocompatibility complex class I phenotype-specificity result in a host immunity that is tightly focused and strain-specific. Immune escape in Theileria is facilitated by genetic diversity in its antigenic determinants, which potentially results in a loss of T cell receptor recognition in its host. In the recent past, several reviews have focused on genetic diversity in the transforming species, Theileriaparva and Theileriaannulata. In contrast, genetic diversity in Theileriaorientalis, a benign non-transforming parasite, which occasionally causes disease outbreaks in cattle, has not been extensively examined. In this review, therefore, we provide an outline of the evolution of Theileria, which includes T. orientalis, and discuss the possible mechanisms generating genetic diversity among parasite populations. Additionally, we discuss the potential implications of a genetically diverse parasite population in the context of Theileria vaccine development.

  13. The Improvement of Maize Pollen In Vitro Germination Method and Its Role in Pollen-mediated Plant Genetic Transformation%玉米花粉体外萌发方法改进及其对花粉介导转基因的作用

    Institute of Scientific and Technical Information of China (English)

    崔贵梅; 孙毅; 郝曜山; 杜建中; 王亦学

    2012-01-01

    The technique of pollen-mediated plant genetic transformation assisted by ultrasonication was patented by the Biotechnology Research Center of the Shanxi Academy of Agricultural Sciences. Here, we further investigated the conditions for sampling, storage and treatment of maize (Zea mays L.) pollen and effects on in vitro germination of pollen with the method. The optimal sucrose concentration for pollen suspension solution differed for maize pollen collected from plants with different flowering time. We determined the storage time and condition of pollen and their effects on pollen germination after ultrasonication. These results would be useful for improving pollen vitality in genetic transformation and verifying the feasibility of this plant transformation technique. We discuss some key steps for conducting in vitro germination of maize pollen and related parameters to increase transformation efficiency of pollen-mediated plant genetic transformation.%超声波处理花粉介导植物基因转化方法由山西省农业科学院生物技术研究中心发明,已被国家知识产权局授予发明专利(专利号ZL 99121152.9).在该专利的基础上,针对玉米(Zea mays)花粉取样、保存和处理条件等因素对其体外萌发的影响进行深入研究,提出了改进玉米花粉体外萌发实验的方法.研究结果表明,在不同时期对开花的玉米进行花粉培养时所需蔗糖溶液的浓度不同;确定了玉米花粉的保存时间、条件及其对超声波处理后花粉萌发率的影响,以提高该转化方法中花粉的活力,并进一步验证了该转基因方法的可靠性;讨论了玉米花粉体外萌发的操作技巧和各因子的参数,对提高花粉介导植物基因转化效率有一定的参考价值.

  14. Radiogenic cell transformation and carcinogenesis

    Science.gov (United States)

    Yang, T. C.; Georgy, K. A.; Mei, M.; Durante, M.; Craise, L. M.

    1995-01-01

    Radiation carcinogenesis is one of the major biological effects considered important in the risk assessment for space travel. Various biological model systems, including both cultured cells and animals, have been found useful for studying the carcinogenic effects of space radiations, which consist of energetic electrons, protons and heavy ions. The development of techniques for studying neoplastic cell transformation in culture has made it possible to examine the cellular and molecular mechanisms of radiation carcinogenesis. Cultured cell systems are thus complementary to animal models. Many investigators have determined the oncogenic effects of ionizing and nonionizing radiation in cultured mammalian cells. One of the cell systems used most often for radiation transformation studies is mouse embryonic cells (C3H10T1/2), which are easy to culture and give good quantitative dose-response curves. Relative biological effectiveness (RBE) for heavy ions with various energies and linear energy transfer (LET) have been obtained with this cell system. Similar RBE and LET relationship was observed by investigators for other cell systems. In addition to RBE measurements, fundamental questions on repair of sub- and potential oncogenic lesions, direct and indirect effect, primary target and lesion, the importance of cell-cell interaction and the role of oncogenes and tumor suppressor genes in radiogenic carcinogenesis have been studied, and interesting results have been found. Recently several human epithelial cell systems have been developed, and ionizing radiation have been shown to transform these cells. Oncogenic transformation of these cells, however, requires a long expression time and/or multiple radiation exposures. Limited experimental data indicate high-LET heavy ions can be more effective than low-LET radiation in inducing cell transformation. Cytogenetic and molecular analyses can be performed with cloned transformants to provide insights into basic genetic

  15. Heterospecific transformation among cyanobacteria.

    OpenAIRE

    Stevens, S. E.; Porter, R D

    1986-01-01

    Heterospecific transformation occurred between cyanobacteria currently classified in either the genus Synechococcus or Synechocystis. Cyanobacterial strains 73109 and 6906 were capable of physiological transformation.

  16. Heterospecific transformation among cyanobacteria.

    OpenAIRE

    Stevens, S E; Porter, R D

    1986-01-01

    Heterospecific transformation occurred between cyanobacteria currently classified in either the genus Synechococcus or Synechocystis. Cyanobacterial strains 73109 and 6906 were capable of physiological transformation.

  17. Transformation of medicinal plants using Agrobacterium tumefaciens.

    Science.gov (United States)

    Bandurska, Katarzyna; Berdowska, Agnieszka; Król, Małgorzata

    2016-12-20

    For many years attempts are made to develop efficient methods for transformation of medicinal plants via Agrobacterium tumefaciens. It is a soil bacteria which possess a natural ability to infect plants in places of injures which results in arise of cancerous growths (crown gall). This is possible thanks a transfer of fragment of Ti plasmid into plant cells and stable integration with a plant genome. Efficiency of medicinal plant transformation depends on many factors for example: Agrobacterium strain, methods and procedures of transformation as well as on plant species, type and age of the explants and regeneration conditions. The main goal of plant transformation is to increase the amount of naturally occurring bioactive compounds and the production of biopharmaceuticals. Genetic plant transformation via bacteria of the genus Agrobacterium is a complex process which requires detailed analysis of incorporated transgene expression and occurs only in the case when the plant cell acquires the ability to regenerate. In many cases, the regeneration efficiency observed in medicinal plants are inefficient after applied transformation procedures. To date there have been attempts of genetic transformation by using A. tumefaciens of medicinal plants belonging to the families: Apocynaceae, Araceae, Araliaceae, Asphodelaceae, Asteraceae, Begoniaceae, Crassulaceae, Fabaceae, Lamiaceae, Linaceae, Papaveraceae, Plantaginaceae, Scrophulariaceae and Solanaceae.

  18. Agrobacterium-mediated transformation of Brachypodium distachyon.

    Science.gov (United States)

    Thole, Vera; Vain, Philippe

    2012-01-01

    Brachypodium distachyon is an attractive genomics and biological model system for grass research. Recently, the complete annotated genome sequence of the diploid line Bd21 has been released. Genetic transformation technologies are critical for the discovery and validation of gene function in Brachypodium. Here, we describe an efficient procedure enabling the Agrobacterium-mediated transformation of a range of diploid and polyploid genotypes of Brachypodium. The procedure relies on the transformation of compact embryogenic calli derived from immature embryos using either chemical selection alone or a combination of chemical and visual screening of transformed tissues and plants. Transformation efficiencies of around 20% can routinely be achieved using this protocol. In the context of the BrachyTAG programme (BrachyTAG.org), this procedure made possible the mass production of Bd21T-DNA mutant plant lines.

  19. 绿色荧光蛋白基因作为报告基因在水稻基因转化中的应用研究%The Applications of GFP as a Reporter Gene in Rice Genetic Transformation

    Institute of Scientific and Technical Information of China (English)

    程在全; WURAY; 等

    2002-01-01

    - fluorescence of rice plants.The GFP was also observed in transformed calli under microscope.The transient expression of GFP helped to demonstrate that the bomba rdment shots of 900psi & 1350 psi make the particles carrying plasmids reach mor e cells than the bombardment shots of 2X 900psi or 2X 1350psi. Our results showe d that this modified GFP gene can be used as a reporter gene for rice genetic tr ansformation and even for other cereal crops such as wheat and corn etc.The resu lt also professes that the MAR sequence can enhance the expression of GFP gene s ignificantly(data was shown in another paper).

  20. General Υ-transformations

    DEFF Research Database (Denmark)

    Barndorff-Nielsen, Ole Eiler; Rosinski, Jan; Thorbjørnsen, Steen

    In this paper we introduce a general class of transformations of (all or most of) the class ML(Rd), of d-dimensional Lévy measures on Rd, into itself. We refer to transformations of this type as Υ transformations (or Upsilon transformations). Closely associated to these are mappings of the set ID...

  1. Negotiated Grammar Transformation

    NARCIS (Netherlands)

    Zaytsev, V.

    2012-01-01

    In this paper, we study controlled adaptability of metamodel transformations. We consider one of the most rigid metamodel transformation formalisms — automated grammar transformation with operator suites, where a transformation script is built in such a way that it is essentially meant to be applica

  2. On Hurwitz transformations

    CERN Document Server

    Hassan, M H; Hassan, M Hage; Kibler, M

    1994-01-01

    A bibliography on the Hurwitz transformations is given. We deal here, with some details, with two particular Hurwitz transformations, viz, the \\grq \\to \\grt Kustaanheimo-Stiefel transformation and its \\grh \\to \\grc compact extension. These transformations are derived in the context of Fock-Bargmann-Schwinger calculus with special emphasis on angular momentum theory.

  3. Generalized Fourier transforms classes

    DEFF Research Database (Denmark)

    Berntsen, Svend; Møller, Steen

    2002-01-01

    The Fourier class of integral transforms with kernels $B(\\omega r)$ has by definition inverse transforms with kernel $B(-\\omega r)$. The space of such transforms is explicitly constructed. A slightly more general class of generalized Fourier transforms are introduced. From the general theory...

  4. A highly efficient method for Agrobacterium mediated transformation ...

    African Journals Online (AJOL)

    ONOS

    2010-08-23

    Aug 23, 2010 ... order to introduce target DNA into the rice cell, of which,. Agrobacterium .... Histochemical assay of -glucuronidase and measurement of GUS activity by ..... Genetic transformation and hybridization: Genomic changes in ...

  5. Genetic Disorders

    Science.gov (United States)

    ... This can cause a medical condition called a genetic disorder. You can inherit a gene mutation from ... during your lifetime. There are three types of genetic disorders: Single-gene disorders, where a mutation affects ...

  6. Genetic modification and genetic determinism.

    Science.gov (United States)

    Resnik, David B; Vorhaus, Daniel B

    2006-06-26

    In this article we examine four objections to the genetic modification of human beings: the freedom argument, the giftedness argument, the authenticity argument, and the uniqueness argument. We then demonstrate that each of these arguments against genetic modification assumes a strong version of genetic determinism. Since these strong deterministic assumptions are false, the arguments against genetic modification, which assume and depend upon these assumptions, are therefore unsound. Serious discussion of the morality of genetic modification, and the development of sound science policy, should be driven by arguments that address the actual consequences of genetic modification for individuals and society, not by ones propped up by false or misleading biological assumptions.

  7. Transforming the Way We Teach Function Transformations

    Science.gov (United States)

    Faulkenberry, Eileen Durand; Faulkenberry, Thomas J.

    2010-01-01

    In this article, the authors discuss "function," a well-defined rule that relates inputs to outputs. They have found that by using the input-output definition of "function," they can examine transformations of functions simply by looking at changes to input or output and the respective changes to the graph. Applying transformations to the input…

  8. Imaging Genetics

    Science.gov (United States)

    Munoz, Karen E.; Hyde, Luke W.; Hariri, Ahmad R.

    2009-01-01

    Imaging genetics is an experimental strategy that integrates molecular genetics and neuroimaging technology to examine biological mechanisms that mediate differences in behavior and the risks for psychiatric disorder. The basic principles in imaging genetics and the development of the field are discussed.

  9. Genetic principles.

    Science.gov (United States)

    Abuelo, D

    1987-01-01

    The author discusses the basic principles of genetics, including the classification of genetic disorders and a consideration of the rules and mechanisms of inheritance. The most common pitfalls in clinical genetic diagnosis are described, with emphasis on the problem of the negative or misleading family history.

  10. Imaging Genetics

    Science.gov (United States)

    Munoz, Karen E.; Hyde, Luke W.; Hariri, Ahmad R.

    2009-01-01

    Imaging genetics is an experimental strategy that integrates molecular genetics and neuroimaging technology to examine biological mechanisms that mediate differences in behavior and the risks for psychiatric disorder. The basic principles in imaging genetics and the development of the field are discussed.

  11. Optimized electroporation-induced transformation in Microcystis aeruginosa PCC7806

    Directory of Open Access Journals (Sweden)

    El Semary, A.

    2010-01-01

    Full Text Available Gene disruption in cyanobacteria is difficult and comprises an obstacle for genetic manipulation. Very few reports tackled this problem but the methods used are usually obscure and hardly reproducible. Here we describe an optimized electroporation-induced transformation in Microcystis aeruginosa PCC7806 where conditions for successful electroporation and transformation are investigated.

  12. On discrete cosine transform

    CERN Document Server

    Zhou, Jianqin

    2011-01-01

    The discrete cosine transform (DCT), introduced by Ahmed, Natarajan and Rao, has been used in many applications of digital signal processing, data compression and information hiding. There are four types of the discrete cosine transform. In simulating the discrete cosine transform, we propose a generalized discrete cosine transform with three parameters, and prove its orthogonality for some new cases. A new type of discrete cosine transform is proposed and its orthogonality is proved. Finally, we propose a generalized discrete W transform with three parameters, and prove its orthogonality for some new cases.

  13. 农杆菌介导的sbe2a基因RNAi载体对玉米遗传转化的研究%Genetic Transformation with RNAi Vector of sbe2a Gene on Maize via Agrobacterium

    Institute of Scientific and Technical Information of China (English)

    关淑艳; 赵丽娜; 刘慧婧; 刘广娜; 王丕武

    2011-01-01

    将淀粉分支酶基因sbe2a的RNAi表达载体转入玉米自交系H99和丹598胚性愈伤组织.探讨了不同菌浓度、不同侵染时间和不同的共培养时间对玉米转化率的影响,确定了最佳转化条件:菌液浓度OD值为0.5~0.7,侵染时间为25 min,共体培养时间为3d.对转化的愈伤组织分化诱导出苗后进行PCR检测,获得了2株阳性植株,初步证明外源基因已经整合到玉米的基因组中.%Starch branch enzyme sbe2a gene of RNAi expression vector was transformed into embryogenic callus in maize inbred lines of H99 and Dan598 via Agrobacterium transformation system. The different concentration of bacteria, the different time of infection and the different time of co-culture on the infection rate of conversion of corn were determined. The results showed that the bacteria concentration (OD600 0. 5 ~ 0.7), infection time (25min)and co-culture time (3d) were optimal. Differentiated transformed callus were detected by PCR, indicating that 2 positive transgenic plants were obtained.

  14. Transformation of undomesticated strains of Bacillus subtilis by protoplast electroporation

    OpenAIRE

    Romero, Diego; Perez-Garcia, Alejandro; Veening, Jan-Willem; Vicente, Antonio; Oscar P. Kuipers; Vicente A.

    2006-01-01

    A rapid method combining the use of protoplasts and electroporation was developed to transform recalcitrant wild strains of Bacillus subtilis. The method described here allows transformation with both replicative and integrative plasmids, as well as with chromosomal DNA, and provides a valuable tool for molecular genetic analysis of interesting Bacillus strains, which are hard to transform by conventional methods. (c) 2006 Elsevier B.V. All rights reserved.

  15. Genetic modification and genetic determinism

    OpenAIRE

    Vorhaus Daniel B; Resnik David B

    2006-01-01

    Abstract In this article we examine four objections to the genetic modification of human beings: the freedom argument, the giftedness argument, the authenticity argument, and the uniqueness argument. We then demonstrate that each of these arguments against genetic modification assumes a strong version of genetic determinism. Since these strong deterministic assumptions are false, the arguments against genetic modification, which assume and depend upon these assumptions, are therefore unsound....

  16. 葡萄抗病毒双价RNAi植物表达载体构建及其对烟草的遗传转化%Construction of binary RNAi expression vector with virus resistance and genetic transformation in tobacco

    Institute of Scientific and Technical Information of China (English)

    田莉莉; 牛良

    2012-01-01

    【Objective】In order to develop new disease-resistant plant materials against two kinds of grape virus,【Method】The binary virus resistant RNAi expression vector was constructed using gateway technology.The conserved sequence of CP(coat protein) gene of Grapevine leafroll virus(GLRaV-3) and Grapevine fanleaf virus(GFLV) were obtained separately via RT-PCR.Then the two conserved sequences were connected to form the interference fragment of GLRaV-GFLV(508 bp) via overlapping PCR.And the fragment was cloned into the vector of pENTR/SD/D to form pENTR/SD/D-GLRaV-GFLV via TOPO cloning.Secondly the ccdB fragment on destination vector pH7GWIWG2(Ⅰ) was replaced by the interference fragment on pENTR/SD/D-GLRaV-GFLV via LR reaction to form the RNAi expression vector of pH7GWIWG2(Ⅰ)-GLRaV-GFLV which contained the two kinds of grape virus gene fragments.Then the vector was transformed into Agrobacterium strain EHA105 by alternate freezing and thawing method.And the RNAi vector was used to transform Nicotiana benthamiana.【Result】The objective fragments were transformed into tobacco successfully by PCR identification.【Conclusion】The Gateway technology-compatible construction of RNAi plant expression vector and tobacco transformation in this study make it possible to induce posttranscriptional gene silencing in the transformed plants and obtain novel double-virus-resistant plant materials.%【目的】为获得兼抗2种葡萄病毒的抗病植物新材料,【方法】利用Gateway技术成功构建了葡萄抗病毒双价RNAi表达载体。先采用RT-PCR分别克隆获得了葡萄卷叶病毒(GLRaV-3)和扇叶病毒(GFLV)外壳蛋白基因保守区段,用重叠延伸PCR方法串联获得了干扰片段GLRaV-GFLV(508 bp);通过TOPO克隆将该片段克隆入载体pENTR/SD/D构建了入门克隆载体pENTR/SD/D-GLRaV-GFLV;再经过LR反应使入门克隆载体pENTR/SD/D-GLRaV-GFLV上的干扰片段GLRaV-GFLV替换目标载体pH7GWIWG2(

  17. A type IV pilus mediates DNA binding during natural transformation in Streptococcus pneumoniae.

    Directory of Open Access Journals (Sweden)

    Raphaël Laurenceau

    Full Text Available Natural genetic transformation is widely distributed in bacteria and generally occurs during a genetically programmed differentiated state called competence. This process promotes genome plasticity and adaptability in Gram-negative and Gram-positive bacteria. Transformation requires the binding and internalization of exogenous DNA, the mechanisms of which are unclear. Here, we report the discovery of a transformation pilus at the surface of competent Streptococcus pneumoniae cells. This Type IV-like pilus, which is primarily composed of the ComGC pilin, is required for transformation. We provide evidence that it directly binds DNA and propose that the transformation pilus is the primary DNA receptor on the bacterial cell during transformation in S. pneumoniae. Being a central component of the transformation apparatus, the transformation pilus enables S. pneumoniae, a major Gram-positive human pathogen, to acquire resistance to antibiotics and to escape vaccines through the binding and incorporation of new genetic material.

  18. Optical source transformations.

    Science.gov (United States)

    Kundtz, N; Roberts, D A; Allen, J; Cummer, S; Smith, D R

    2008-12-22

    Transformation optics is a recently appreciated methodology for the design of complex media that control the propagation of electromagnetic and other types of waves. The transformation optical technique involves the use of coordinate transformations applied to some region of space, providing a conceptual means to redirect the flow of waves. Successfully designed devices to date have made use of transformations acting on passive space only; however, the technique can also be applied when source distributions (e.g., current and charge) are included within the space being transformed. In this paper we present examples of source transformations that illustrate the potential of these expanded transformation optical methods. In particular, using finite-element full-wave simulations, we confirm the restoration of dipole radiation patterns from both a distorted 'pin-wheel' antenna and a bent dipole partially occluded by a cylindrical scatterer. We propose the technique of source transformations as a powerful approach for antenna design, especially in relation to conformal antennas.

  19. 28-Channel rotary transformer

    Science.gov (United States)

    Mclyman, W. T.

    1981-01-01

    Transformer transmits power and digital data across rotating interface. Array has many parallel data channels, each with potential l megabaud data rate. Ferrite-cored transformers are spaced along rotor; airgap between them reduces crosstalk.

  20. Steerable Discrete Cosine Transform

    OpenAIRE

    Fracastoro, Giulia; Fosson, Sophie; Magli, Enrico

    2017-01-01

    In image compression, classical block-based separable transforms tend to be inefficient when image blocks contain arbitrarily shaped discontinuities. For this reason, transforms incorporating directional information are an appealing alternative. In this paper, we propose a new approach to this problem, namely, a discrete cosine transform (DCT) that can be steered in any chosen direction. Such transform, called steerable DCT (SDCT), allows to rotate in a flexible way pairs of basis vectors, an...

  1. Entropy of Baker's Transformation

    Institute of Scientific and Technical Information of China (English)

    栾长福

    2003-01-01

    Four theorems about four different kinds of entropies for Baker's transformation are presented. The Kolmogorov entropy of Baker's transformation is sensitive to the initial flips by the time. The topological entropy of Baker's transformation is found to be log k. The conditions for the state of Baker's transformation to be forbidden are also derived. The relations among the Shanonn, Kolmogorov, topological and Boltzmann entropies are discussed in details.

  2. Fourier and Laplace Transforms

    NARCIS (Netherlands)

    Beerends, R.J.; Morsche, ter H.G.; Berg, van den J.C.

    2003-01-01

    This textbook presents in a unified manner the fundamentals of both continuous and discrete versions of the Fourier and Laplace transforms. These transforms play an important role in the analysis of all kinds of physical phenomena. As a link between the various applications of these transforms the a

  3. SELECTION OF POWER TRANSFORMERS

    OpenAIRE

    A. A. Gonchar

    2005-01-01

    The paper reveals that while loading transformers from idle running till nominal value it is preferable to apply transformers with less power losses in steel because total losses of active power in the indicated range are less than in transformers with larger losses in steel.

  4. Genomics, proteomics, and genetics of leptospira.

    Science.gov (United States)

    Picardeau, Mathieu

    2015-01-01

    Recent advances in molecular genetics, such as the ability to construct defined mutants, have allowed the study of virulence factors and more generally the biology in Leptospira. However, pathogenic leptospires remain much less easily transformable than the saprophyte L. biflexa and further development and improvement of genetic tools are required. Here, we review tools that have been used to genetically manipulate Leptospira. We also describe the major advances achieved in both genomics and postgenomics technologies, including transcriptomics and proteomics.

  5. Chapter VIII. Contributions of propagation techniques and genetic modification to breeding - genetic engineering for disease resistance

    Science.gov (United States)

    Genetic engineering offers an opportunity to develop flower bulb crops with resistance to fungal, viral, and bacterial pathogens. Several of the flower bulb crops, Lilium spp., Gladiolus, Zantedeschia, Muscari, Hyacinthus, Narcissus, Ornithogalum, Iris, and Alstroemeria, have been transformed with t...

  6. Innovations in transforming organizations.

    Science.gov (United States)

    Simpson, Roy L

    2009-01-01

    Transformations take as many forms as the organizations they change. However, the success of healthcare-based transformations requires significant cultural shifts and operational changes, many of which mirror the tenets of classic transformer W. Edward Deming. Attaining Magnet hospital status and the integration of the e-health record into nursing school curricula are 2 examples of how healthcare leaders have championed a series of changes that produced sweeping transformations. Success is likely to elude healthcare leaders who mount massive transformations, while those who focus on making seemingly small, interconnected changes are more likely to see the results they envision over time.

  7. Evaluación y selección de un protocolo vía Agrobacterium para la incorporación de resistencia al cogollero en la variedad de tomate Unapal-Arreboles Evaluation and selection of a protocol for Agrobacterium-mediated genetic transformation of tomato variety Unapal-Arreboles for resistance to budworm

    Directory of Open Access Journals (Sweden)

    Hernando Ramírez

    2009-04-01

    Full Text Available Se evaluó y seleccionó una metodología para la transformación genética de la variedad de tomate UNAPAL-Arreboles con el gen cry1Ab para la incorporación de resistencia al cogollero (Tuta absoluta, utilizando el sistema de Agrobacterium. Se regeneraron 59 plantas transgénicas a partir de 3.200 explantes (1.84%. La integración estable, expresión y herencia de los genes nptII y gus-intrón, se demostraron mediante análisis histoquímico y molecular en los clones To28, To33 y To47 y en la correspondiente generación T1. Sin embargo, los análisis molecular e inmunológico indicaron ausencia del gen cry1Ab sugiriendo que la secuencia de este gen se puede haber modificado.A plant transformation methodology was selected and evaluated to incorporate the cry1Ab gene by Agrobacterium-mediate genetic transformation into tomato variety UNAPAL-Arreboles for resistance to budworm (Tuta absoluta. A total of 59 transgenic plants were regenerated from 3.200 explants (1.84%. Histochemical gus assay and molecular analysis of three independent events To28, To33 and To47 and corresponding T1 derived generations, demonstrate the stable integration, expression and inheritance of the nptII and gus-intron genes. However, the molecular and immunological analysis of these same clones, indicate that the cry1Ab gene is not present in the transformed plants, suggesting that the sequence of this gene may be modified as result of possible recombinant events.

  8. Genetic barcodes

    Energy Technology Data Exchange (ETDEWEB)

    Weier, Heinz -Ulrich G

    2015-08-04

    Herein are described multicolor FISH probe sets termed "genetic barcodes" targeting several cancer or disease-related loci to assess gene rearrangements and copy number changes in tumor cells. Two, three or more different fluorophores are used to detect the genetic barcode sections thus permitting unique labeling and multilocus analysis in individual cell nuclei. Gene specific barcodes can be generated and combined to provide both numerical and structural genetic information for these and other pertinent disease associated genes.

  9. 转基因豆粕对崂山奶山羊生长性能、肌肉营养成分及组织器官中外源基因转移的影响%Effects of Genetically Modified Soybean Meal on Growth Performance, Meat Nutrient Composition, and Exogenous Gene Transformation to Tissues and Organs of Laoshan Dairy Goats

    Institute of Scientific and Technical Information of China (English)

    刘斌; 秦志华; 黄娟; 张廷荣; 刘文达; 王述柏

    2014-01-01

    In order to analyze the effects of genetically modified soybean meal on growth performance, meat nutrient composition, and exogenous gene transformation to tissues and organs of Laoshan dairy goats, thirty 50-day-old Laoshan dairy goats were randomly divided into two groups with three replicates in each group and five goats per replicate. Goats in different groups were fed concentrates containing non-genetically modified soybean meal and genetically modified soybean meal ( MON89788 and GTS40-3-2 ) , respectively, and were fed the same roughage ( Chinese wildrye) . The experiment lasted for 150 days. The results showed as fol-lows: average daily feed intake, average daily gain and feed to gain ratio in the two groups were not signifi-cantly different (P>0. 05); the contents of dry matter, ether extract, crude protein, ash, calcium and phos-phorus in muscle of dairy goats were not significantly different either ( P >0 . 05 ) . Moreover, no gene rag-ments of genetically modified soybean meal were detected from muscle, liver, pancreas, kidney, spleen, thy-mus, lung, heart, abomasum and small intestine of dairy goats by real-time quantification PCR. The results suggest that genetically modified soybean meal ( MON89788 and GTS40-3-2 ) had no significant effects on growth performance and muscle quality of Laoshan dairy goats, and no genetic horizontal transformation is ob-served in tissues and organs.%本试验旨在研究转基因豆粕对崂山奶山羊生长性能、肌肉营养成分及组织器官中外源基因转移的影响。选取30只50日龄崂山奶山羊随机分为2组,每组3个重复,每个重复5只。精饲料分别以非转基因豆粕和转基因豆粕( MON89788和GTS40-3-2)配制,粗饲料均为干草。试验期150 d。结果表明:2组奶山羊平均日采食量、平均日增重和料重比均无显著差异( P >0.05);奶山羊肌肉的干物质、粗脂肪、粗蛋白质、粗灰分、钙、磷含量2组间亦均无显著差异( P>0

  10. Wavelets associated with Hankel transform and their Weyl transforms

    Institute of Scientific and Technical Information of China (English)

    PENG; Lizhong; MA; Ruiqin

    2004-01-01

    The Hankel transform is an important transform. In this paper, westudy the wavelets associated with the Hankel transform, thendefine the Weyl transform of the wavelets. We give criteria of itsboundedness and compactness on the Lp-spaces.

  11. Efficient production of transgenic melon via Agrobacterium-mediated transformation.

    Science.gov (United States)

    Bezirganoglu, I; Hwang, S Y; Shaw, J F; Fang, T J

    2014-04-25

    Oriental melon (Cucumis melo L. var. makuwa) is an important fruit for human consumption. However, this plant species is one of the most recalcitrant to genetic transformation. The lack of an efficient in vitro system limits the development of a reproducible genetic transformation protocol for Oriental melon. In this study, an efficient transgenic production method for Agrobacterium-mediated transformation using cotyledon explants of Oriental melon was developed. Cotyledon explants were pre-cultivated for two days in the dark, and the optimal conditions for transformation of melon were determined to be a bacteria concentration of OD600 0.6, inoculation for 30 min, and two days of co-cultivation. Transgenic melon plants were produced from kanamycin-resistant shoots. A total of 11 independent transgenic plants were regenerated with a transformation efficiency of 0.8% of the inoculated explants. The transgenic plants were phenotypically normal and fully fertile, which might be a consequence of the co-cultivation time.

  12. Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum

    Energy Technology Data Exchange (ETDEWEB)

    Guss, Adam M [ORNL; Olson, Daniel G. [Thayer School of Engineering at Dartmouth; Caiazza, Nicky [Mascoma Corporation; Lynd, Lee R [Thayer School of Engineering at Dartmouth

    2012-01-01

    BACKGROUND: Industrial production of biofuels and other products by cellulolytic microorganisms is of interest but hindered by the nascent state of genetic tools. Although a genetic system for Clostridium thermocellum DSM1313 has recently been developed, available methods achieve relatively low efficiency and similar plasmids can transform C. thermocellum at dramatically different efficiencies. RESULTS: We report an increase in transformation efficiency of C. thermocellum for a variety of plasmids by using DNA that has been methylated by Escherichia coli Dam but not Dcm methylases. When isolated from a dam+ dcm+ E. coli strain, pAMG206 transforms C. thermocellum 100-fold better than the similar plasmid pAMG205, which contains an additional Dcm methylation site in the pyrF gene. Upon removal of Dcm methylation, transformation with pAMG206 showed a four- to seven-fold increase in efficiency; however, transformation efficiency of pAMG205 increased 500-fold. Removal of the Dcm methylation site from the pAM205 pyrF gene via silent mutation resulted in increased transformation efficiencies equivalent to that of pAMG206. Upon proper methylation, transformation efficiency of plasmids bearing the pMK3 and pB6A origins of replication increased ca. three orders of magnitude. CONCLUSION: E. coli Dcm methylation decreases transformation efficiency in C. thermocellum DSM1313. The use of properly methylated plasmid DNA should facilitate genetic manipulation of this industrially relevant bacterium.

  13. Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum

    Directory of Open Access Journals (Sweden)

    Guss Adam M

    2012-05-01

    Full Text Available Abstract Background Industrial production of biofuels and other products by cellulolytic microorganisms is of interest but hindered by the nascent state of genetic tools. Although a genetic system for Clostridium thermocellum DSM1313 has recently been developed, available methods achieve relatively low efficiency and similar plasmids can transform C. thermocellum at dramatically different efficiencies. Results We report an increase in transformation efficiency of C. thermocellum for a variety of plasmids by using DNA that has been methylated by Escherichia coli Dam but not Dcm methylases. When isolated from a dam+dcm+E. coli strain, pAMG206 transforms C. thermocellum 100-fold better than the similar plasmid pAMG205, which contains an additional Dcm methylation site in the pyrF gene. Upon removal of Dcm methylation, transformation with pAMG206 showed a four- to seven-fold increase in efficiency; however, transformation efficiency of pAMG205 increased 500-fold. Removal of the Dcm methylation site from the pAMG205 pyrF gene via silent mutation resulted in increased transformation efficiencies equivalent to that of pAMG206. Upon proper methylation, transformation efficiency of plasmids bearing the pMK3 and pB6A origins of replication increased ca. three orders of magnitude. Conclusions E. coli Dcm methylation decreases transformation efficiency in C. thermocellum DSM1313. The use of properly methylated plasmid DNA should facilitate genetic manipulation of this industrially relevant bacterium.

  14. Genetic modification and genetic determinism

    Directory of Open Access Journals (Sweden)

    Vorhaus Daniel B

    2006-06-01

    Full Text Available Abstract In this article we examine four objections to the genetic modification of human beings: the freedom argument, the giftedness argument, the authenticity argument, and the uniqueness argument. We then demonstrate that each of these arguments against genetic modification assumes a strong version of genetic determinism. Since these strong deterministic assumptions are false, the arguments against genetic modification, which assume and depend upon these assumptions, are therefore unsound. Serious discussion of the morality of genetic modification, and the development of sound science policy, should be driven by arguments that address the actual consequences of genetic modification for individuals and society, not by ones propped up by false or misleading biological assumptions.

  15. 基于多目标遗传算法的水陆两栖可变形机器人结构参数设计方法%Mechanism-parameters Design Method of an Amphibious Transformable Robot Based on Multi-objective Genetic Algorithm

    Institute of Scientific and Technical Information of China (English)

    李楠; 王明辉; 马书根; 李斌; 王越超

    2012-01-01

    As a new style of the mobile robot, the amphibious transformable robot can not only perform reconfiguration but also implement tasks in amphibious environment. For the mechanism design of the robot, the parameter of the mechanism takes influence on the robot's performance in the task environment. To implement the performance optimization in the complex environment and task, a mechanism-parameters design method of an amphibious transformable robot based on multi-objective genetic algorithm is proposed. Based on the kinematics and dynamic analysis of the robot, the multi-objective optimization problem of the mechanism parameters design is established on the mapping relationships between the performance indexes and the mechanism parameters. The non-dominated sorting genetic algorithm II (NSGA-II) is adopted to solve this optimization problem and get the Pareto optimization. A solution of optimization problem of the mechanism parameters is extracted through the Pareto optimization based on the optimizing method of combination weighting of multi-attribute decision-making, and then the result is used to direct the mechanism design of the amphibious transformable robot, Amoeba-II. The experiment for the maneuverability of Amoeba-II in the amphibious environment is performed to certify the validity and applicability of the mechanism-parameters design method of amphibious transformable robot based on multi-objective genetic algorithm.%水陆两栖可变形机器人是一种兼具变形能力与两栖环境适应能力的新型移动机器人.在其机构设计中,结构参数直接影响该机器人在任务环境中的各项机动性能.针对水陆两栖可变形机器人工作环境复杂性和任务多变性,提出一种基于多目标遗传算法的机器人结构参数设计方法,以得到该型机器人在两栖环境中的最优的综合性能.在水陆两栖可变形机器人陆地环境和水环境中运动学和动力学模型基础上,建立两栖环境中机

  16. CLONING AND GENETIC TRANSFORMATION OF fw2.2 GENE IN PEANUT%花生fw2.2基因的克隆及遗传转化

    Institute of Scientific and Technical Information of China (English)

    刘强; 王晶珊; 乔利仙; 赵春梅; 隋炯明

    2011-01-01

    fw2.2基因是影响番茄果重的一个重要数量性状基因,在心皮的细胞分裂中起负调控作用。本研究以番茄fw2.2基因的序列为探针,从花生EST数据库中筛选同源序列。根据花生的EST拼接序列设计引物对花生进行扩增,目标产物测序后,将推测的氨基酸序列与其他植物fw2.2基因编码的氨基酸序列进行比对,同源性为34.78%~66.85%。半定量RT-PCR结果表明,该基因在野生种和栽培种中的表达存在差异。将fw2.2基因连接到植物表达载体,通过农杆菌介导法转化花生品种花育23号,获得了12个PCR阳性的独立转化子。%fw2.2 gene is an important quantative trait locus(QTL) affecting fruit weight in tomato,and can reduce cell division in carpels as a negative regulator.In this experiment,fw2.2 gene in tomato was prepared as probe to screen homologous sequence from peanut EST database.Then,according to the joint EST sequence,a pair of primer was designed to amplified the fw2.2 gene of peanut.Compared with other plants,the similarity range of amino acid sequence was between 34.78% and 66.85%.Expression difference was detected between wild and cultivated materials by semi-quantitative RT-PCR.After transformation mediated by Agrobacterium tumefaciens,12 independent transformants of peanut cultivar Huayu 23 were identificated by PCR amplification.

  17. Genetic Engineering

    Science.gov (United States)

    Phillips, John

    1973-01-01

    Presents a review of genetic engineering, in which the genotypes of plants and animals (including human genotypes) may be manipulated for the benefit of the human species. Discusses associated problems and solutions and provides an extensive bibliography of literature relating to genetic engineering. (JR)

  18. Genetic Counseling

    Science.gov (United States)

    ... for certain types of genetic conditions (such as Down syndrome) in the baby if mother-to-be is 35 years of age or more, or is concerned at any age about her chances of having a child with a genetic condition To learn about the ...

  19. 大豆铁蛋白基因GmFerritin在玉米中的遗传转化%Genetic transformation of GmFerritin in maize (Zea mays L.)

    Institute of Scientific and Technical Information of China (English)

    李晓丽; 王建军; 崔瑞洁; 谢传晓; 段运平

    2012-01-01

    The plant expression vector pCUB-Zein::ferritin-35s::bar was constructed by In-Fusion kit. The soybean ferritin gene was transferred into the shoot apical point of maize inbred line of maize inbred line Qi319 by Agrobacterium tumefacien mediated transformation. The soybean ferritin gene was expressed under the control of a maize seed storage protein prolamin promoter 15kD p-Ze/w. 272 transgenic plants were obtained after select ing with glufosinate and 108 of them were confirmed positive by PCR amplification. The frequency of transfor mation reached about 3.6%. This study showed that the soybean ferritin gene was integrated into maize genome.%使用In-Fusion试剂盒构建表达载体pCUB-Zein::ferritin-3 5s::bar,以玉米自交系齐319茎尖分生组织为受体,采用农杆菌介导法,将玉米胚乳特异启动子基因15 kD β-Zein驱动的大豆铁蛋白ferritin基因转入玉米,共筛选出272株除草剂(草铵膦)抗性植株,其中108株PCR检测呈阳性,转化率达3.6%,初步判断Zein:ferritin 基因已转入玉米基因组中.

  20. Genetic Romanticism

    DEFF Research Database (Denmark)

    Tupasela, Aaro

    2016-01-01

    . This article compares and contrasts the work of two doctors in Finland, Elias Lönnrot and Reijo Norio, working over a century and a half apart, to examine the ways in which they have contributed to the formation of national identity and unity. The notion of genetic romanticism is introduced as a term...... to complement the notion of national romanticism that has been used to describe the ways in which nineteenth-century scholars sought to create and deploy common traditions for national-romantic purposes. Unlike national romanticism, however, strategies of genetic romanticism rely on the study of genetic...... inheritance as a way to unify populations within politically and geographically bounded areas. Thus, new genetics have contributed to the development of genetic romanticisms, whereby populations (human, plant, and animal) can be delineated and mobilized through scientific and medical practices to represent...

  1. Genetically Engineering Entomopathogenic Fungi.

    Science.gov (United States)

    Zhao, H; Lovett, B; Fang, W

    2016-01-01

    Entomopathogenic fungi have been developed as environmentally friendly alternatives to chemical insecticides in biocontrol programs for agricultural pests and vectors of disease. However, mycoinsecticides currently have a small market share due to low virulence and inconsistencies in their performance. Genetic engineering has made it possible to significantly improve the virulence of fungi and their tolerance to adverse conditions. Virulence enhancement has been achieved by engineering fungi to express insect proteins and insecticidal proteins/peptides from insect predators and other insect pathogens, or by overexpressing the pathogen's own genes. Importantly, protein engineering can be used to mix and match functional domains from diverse genes sourced from entomopathogenic fungi and other organisms, producing insecticidal proteins with novel characteristics. Fungal tolerance to abiotic stresses, especially UV radiation, has been greatly improved by introducing into entomopathogens a photoreactivation system from an archaean and pigment synthesis pathways from nonentomopathogenic fungi. Conversely, gene knockout strategies have produced strains with reduced ecological fitness as recipients for genetic engineering to improve virulence; the resulting strains are hypervirulent, but will not persist in the environment. Coupled with their natural insect specificity, safety concerns can also be mitigated by using safe effector proteins with selection marker genes removed after transformation. With the increasing public concern over the continued use of synthetic chemical insecticides and growing public acceptance of genetically modified organisms, new types of biological insecticides produced by genetic engineering offer a range of environmentally friendly options for cost-effective control of insect pests. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Genetically modified yeast species, and fermentation processes using genetically modified yeast

    Science.gov (United States)

    Rajgarhia, Vineet; Koivuranta, Kari; Penttila, Merja; Ilmen, Marja; Suominen, Pirkko; Aristidou, Aristos; Miller, Christopher Kenneth; Olson, Stacey; Ruohonen, Laura

    2013-05-14

    Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

  3. Genetically modified yeast species, and fermentation processes using genetically modified yeast

    Energy Technology Data Exchange (ETDEWEB)

    Rajgarhia, Vineet; Koivuranta, Kari; Penttila, Merja; Ilmen, Marja; Suominen, Pirkko; Aristidou, Aristos; Miller, Christopher Kenneth; Olson, Stacey; Ruohonen, Laura

    2016-08-09

    Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

  4. Genetically modified yeast species, and fermentation processes using genetically modified yeast

    Energy Technology Data Exchange (ETDEWEB)

    Rajgarhia, Vineet; Koivuranta, Kari; Penttila, Merja; Ilmen, Marja; Suominen, Pirkko; Aristidou, Aristos; Miller, Christopher Kenneth; Olson, Stacey; Ruohonen, Laura

    2017-09-12

    Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific or specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

  5. Genetically modified yeast species, and fermentation processes using genetically modified yeast

    Energy Technology Data Exchange (ETDEWEB)

    Rajgarhia, Vineet [Kingsport, TN; Koivuranta, Kari [Helsinki, FI; Penttila, Merja [Helsinki, FI; Ilmen, Marja [Helsinki, FI; Suominen, Pirkko [Maple Grove, MN; Aristidou, Aristos [Maple Grove, MN; Miller, Christopher Kenneth [Cottage Grove, MN; Olson, Stacey [St. Bonifacius, MN; Ruohonen, Laura [Helsinki, FI

    2014-01-07

    Yeast cells are transformed with an exogenous xylose isomerase gene. Additional genetic modifications enhance the ability of the transformed cells to ferment xylose to ethanol or other desired fermentation products. Those modifications include deletion of non-specific aldose reductase gene(s), deletion of xylitol dehydrogenase gene(s) and/or overexpression of xylulokinase.

  6. Genetic testing in cardiovascular diseases.

    Science.gov (United States)

    Arndt, Anne-Karin; MacRae, Calum A

    2014-05-01

    The review is designed to outline the major developments in genetic testing in the cardiovascular arena in the past year or so. This is an exciting time in genetic testing as whole exome and whole genome approaches finally reach the clinic. These new approaches offer insight into disease causation in families in which this might previously have been inaccessible, and also bring a wide range of interpretative challenges. Among the most significant recent findings has been the extent of physiologic rare coding variation in the human genome. New disease genes have been identified through whole exome studies in neonatal arrhythmia, congenital heart disease and coronary artery disease that were simply inaccessible with other techniques. This has not only shed light on the challenges of genetic testing at this scale, but has also sharply defined the limits of prior gene-panel focused testing. As novel therapies targeting specific genetic subsets of disease become available, genetic testing will become a part of routine clinical care. The pace of change in sequencing technologies has begun to transform clinical medicine, and cardiovascular disease is no exception. The complexity of such studies emphasizes the importance of real-time communication between the genetics laboratory and genetically informed clinicians. New efforts in data and knowledge management will be central to the continued advancement of genetic testing.

  7. Evidence for stable transformation of wheat by floraldip in Agrobacterium tumefaciens

    Science.gov (United States)

    Hexaploid wheat is one of the world’s most important staple crops but genetic transformation is still challenging. We have developed a floral transformation protocol that does not utilize tissue culture. Three T-DNA wheat transformants have been produced in the germplasm line, Crocus, using this p...

  8. Steerable Discrete Cosine Transform

    Science.gov (United States)

    Fracastoro, Giulia; Fosson, Sophie M.; Magli, Enrico

    2017-01-01

    In image compression, classical block-based separable transforms tend to be inefficient when image blocks contain arbitrarily shaped discontinuities. For this reason, transforms incorporating directional information are an appealing alternative. In this paper, we propose a new approach to this problem, namely a discrete cosine transform (DCT) that can be steered in any chosen direction. Such transform, called steerable DCT (SDCT), allows to rotate in a flexible way pairs of basis vectors, and enables precise matching of directionality in each image block, achieving improved coding efficiency. The optimal rotation angles for SDCT can be represented as solution of a suitable rate-distortion (RD) problem. We propose iterative methods to search such solution, and we develop a fully fledged image encoder to practically compare our techniques with other competing transforms. Analytical and numerical results prove that SDCT outperforms both DCT and state-of-the-art directional transforms.

  9. Generalizing the Lorentz transformations

    CERN Document Server

    Chappell, James M; Iannella, Nicolangelo; Hartnett, John G; Iqbal, Azhar; Abbott, Derek

    2016-01-01

    In this paper we develop a framework allowing a natural extension of the Lorentz transformations. To begin, we show that by expanding conventional four-dimensional spacetime to eight-dimensions that a natural generalization is indeed obtained. We then find with these generalized coordinate transformations acting on Maxwell's equations that the electromagnetic field transformations are nevertheless unchanged. We find further, that if we assume the absence of magnetic monopoles, in accordance with Maxwell's theory, our generalized transformations are then restricted to be the conventional ones. While the conventional Lorentz transformations are indeed recovered from our framework, we nevertheless provide a new perspective into why the Lorentz transformations are constrained to be the conventional ones. Also, this generalized framework may assist in explaining several unresolved questions in electromagnetism as well as to be able to describe quasi magnetic monopoles found in spin-ice systems.

  10. Transformative environmental governance

    Science.gov (United States)

    Chaffin, Brian C.; Garmestani, Ahjond S.; Gunderson, Lance H.; Harm Benson, Melinda; Angeler, David G.; Arnold, Craig Anthony (Tony); Cosens, Barbara; Kundis Craig, Robin; Ruhl, J.B.; Allen, Craig R.

    2016-01-01

    Transformative governance is an approach to environmental governance that has the capacity to respond to, manage, and trigger regime shifts in coupled social-ecological systems (SESs) at multiple scales. The goal of transformative governance is to actively shift degraded SESs to alternative, more desirable, or more functional regimes by altering the structures and processes that define the system. Transformative governance is rooted in ecological theories to explain cross-scale dynamics in complex systems, as well as social theories of change, innovation, and technological transformation. Similar to adaptive governance, transformative governance involves a broad set of governance components, but requires additional capacity to foster new social-ecological regimes including increased risk tolerance, significant systemic investment, and restructured economies and power relations. Transformative governance has the potential to actively respond to regime shifts triggered by climate change, and thus future research should focus on identifying system drivers and leading indicators associated with social-ecological thresholds.

  11. Transformation of coals at the heat treatment

    Energy Technology Data Exchange (ETDEWEB)

    S.A. Aipshtein; V.I. Minaev; O.V. Barabanova; V.A. Novikova [Moscow State University of Mining (MSMU), Moscow (Russian Federation)

    2007-07-01

    The coals (from Kuznetsk and Donetsk basins) were selected with the following criteria: a) different rank; b) different genetic types. The substantial and chemical compositions of the coals were determined. Experimental works on coal heat treatment were carried out at temperatures 423K, 473K, 523K, 573K and different times of processing in the conditions excluding oxidation of coals. It was shown, that the prevailing contribution of destruction or cross-linking in the general process of coal transformation at the heat treatment depends on temperature and time of thermal treatment. It was shown that the depth and direction of coals transformations at the heat treatment essentially depended on the genetic type of coals. 7 refs., 3 figs., 1 tab.

  12. Steerable Discrete Cosine Transform

    OpenAIRE

    Magli, Enrico; Fracastoro, Giulia

    2015-01-01

    Block-based separable transforms tend to be inefficient when blocks contain arbitrarily shaped discontinuities. For this reason, transforms incorporating directional information are an appealing alternative. In this paper, we propose a new approach to this problem, designing a new transform that can be steered in any chosen direction and that is defined in a rigorous mathematical way. This new steerable DCT allows to rotate in a flexible way pairs of basis vectors, enabling precise matching o...

  13. The Discrete Wavelet Transform

    Science.gov (United States)

    1991-06-01

    focuses on bringing together two separately motivated implementations of the wavelet transform , the algorithm a trous and Mallat’s multiresolution...decomposition. These algorithms are special cases of a single filter bank structure, the discrete wavelet transform , the behavior of which is governed by...nonorthogonal multiresolution algorithm for which the discrete wavelet transform is exact. Moreover, we show that the commonly used Lagrange a trous

  14. Isolation and Homologous Genetic Transformation of DFR in Saussurea involucrata Kar.et Kir.%新疆雪莲DFR基因的分离及同源遗传转化

    Institute of Scientific and Technical Information of China (English)

    覃建兵; 唐亚萍; 曾卫军

    2012-01-01

    二氢黄酮醇4-还原酶(dihydroflavonol 4-reductase,DFR)是植物重要的次生代谢产物花青素生物合成途径中的关键酶.运用RT-PCR和RACE技术从新疆雪莲(Saussurea involucrata Kar.et Kir.)中克隆得到DFR基因(GenBank登录号为JN092126).DFR基因的cDNA全长序列含有1个1 029 bp的开放阅读框(ORF),编码343个氨基酸,该基因推断的蛋白与水母雪莲DFR基因推断的蛋白高度同源,相似性达到92%;以不同物种中DFR氨基酸序列进行比对分析,推断的蛋白含有与NADPH特异结合的结构域.将该基因运用农杆菌介导的叶片转化法进行同源转化,将含有转DFR基因的愈伤组织进行悬浮培养,紫外分光光度法测定愈伤组织的总黄酮含量,结果表明转基因愈伤组织的总黄酮含量明显高于非转基因愈伤组织的含量.该研究为提高新疆雪莲药用化学成分黄酮类物质及实现新疆雪莲花青素的人工生物合成的研究奠定基础,对解决天山雪莲资源匮乏提供参考.%The dihydroflavonol-4-reductase (DFR) is an important enzyme in plant secondary metabolites and catalyzes the key step in the biosynthesis of anthocyanins. DFR gene ( GenBank Accession JN092126) was cloned from Saussurea involucrata Kar. et Kir. by RT-PCR and RACE. The cDNA sequence of DFR was consisted of 1 029 bp open reading frame ( ORF) encoding 343 amino acid, the deduced DFR protein has high homolo-gy with S. medusa (92% identity). Homology analysis showed that deduced DFR protein has the special domain regioselectivity towards NADPH. DFR gene of S. involucrata was under the control of the cauliflower mosaic virus (CaMV) 35S promoter, homologous transformation was conducted using an Agrobacter-ium rihizogenes-mediated transformation system. The results of UV spectrophotometry showed that Sinvolucrata callus after suspension culture had an obviously higher average content of total flavonoids than non-transgenic callus. This study will improve the

  15. Transformação genética e aplicação de glifosato na microbiota do solo, fixação biológica de nitrogênio, qualidade e segurança de grãos de soja geneticamente modificada Genetic transformation and the use glyphosate on soil microbial, biological nitrogen fixation, quality and safety of genetically modified soybean

    Directory of Open Access Journals (Sweden)

    Giani Mariza Bärwald Böhm

    2010-02-01

    safety of GM RR soybean with or without glyphosate application were collected and investigated. In general, the researches investigated showed some similar characteristics: a few variables were studied; b simulations in green house or in vitro are used to infer possible response on the field. Although scarce, the researches denoted that the genetic transformation did not affect soil microorganisms, biological nitrogen fixation and composition of grains. The factor which could affect these variables is the glyphosate application in weed control, thus the risk of damage to the environment and the safety of products derived from this raw materialare due to the use of the herbicide during cultivation, and not to the genetic transformation. Key words: Transgenic soybean, soil microorganisms, herbicides residue, isoflavones.

  16. Phase transformation and diffusion

    CERN Document Server

    Kale, G B; Dey, G K

    2008-01-01

    Given that the basic purpose of all research in materials science and technology is to tailor the properties of materials to suit specific applications, phase transformations are the natural key to the fine-tuning of the structural, mechanical and corrosion properties. A basic understanding of the kinetics and mechanisms of phase transformation is therefore of vital importance. Apart from a few cases involving crystallographic martensitic transformations, all phase transformations are mediated by diffusion. Thus, proper control and understanding of the process of diffusion during nucleation, g

  17. Distributed photovoltaic grid transformers

    CERN Document Server

    Shertukde, Hemchandra Madhusudan

    2014-01-01

    The demand for alternative energy sources fuels the need for electric power and controls engineers to possess a practical understanding of transformers suitable for solar energy. Meeting that need, Distributed Photovoltaic Grid Transformers begins by explaining the basic theory behind transformers in the solar power arena, and then progresses to describe the development, manufacture, and sale of distributed photovoltaic (PV) grid transformers, which help boost the electric DC voltage (generally at 30 volts) harnessed by a PV panel to a higher level (generally at 115 volts or higher) once it is

  18. A DC Transformer Project

    Data.gov (United States)

    National Aeronautics and Space Administration — During the period of this project three different possible DC transformer concepts were proposed, theoretically modeled, and then experimentally tested with the...

  19. Martensitic transformations; Martensite hentai

    Energy Technology Data Exchange (ETDEWEB)

    Otsuka, K. [University of Tsukuba, Tsukuba (Japan)

    1997-09-20

    This paper explains the recent studies on martensitic transformation (M transformation). The classical theory on nucleation in M transformation gives non-realistic activation energy as large as 10{sup 4}eV for Fe system. Although various theoretical and experimental approaches have been attempted, a universal theory is not yet established. The {beta}{sub 1}(DO{sub 3})-{gamma}{sub 1} transformation of Cu- Al-Ni alloy was resolved as lattice-invariable deformation by introducing the second kind twin. Various subsequent comparative studies for alloy systems between a phenomenology and experimental results showed the validity of a phenomenology due to lattice-invariable deformation. In the thermodynamics of M transformation, it has been recognized that the mechanism of transformation is entirely different between athermal and isothermal transformations, however, a statistical thermodynamic model was proposed for dealing with these transformations integrally. The study on intelligent materials is under active investigation from the viewpoint of application of M transformation. 44 refs.

  20. Program Transformation in HATS

    Energy Technology Data Exchange (ETDEWEB)

    Winter, V.L.

    1999-02-24

    HATS is a general purpose syntax derivation tree based transformation system in which transformation sequences are described in special purpose language. A powerful feature of this language is that unification is an explicit operation. By making unification explicit, an elegant framework arises in which to express complex application conditions which in turn enables refined control strategies to be realized. This paper gives an overview of HATS, focusing especially on the framework provided by the transformation language and its potential with respect to control and general purpose transformation.

  1. The convolution transform

    CERN Document Server

    Hirschman, Isidore Isaac

    2005-01-01

    In studies of general operators of the same nature, general convolution transforms are immediately encountered as the objects of inversion. The relation between differential operators and integral transforms is the basic theme of this work, which is geared toward upper-level undergraduates and graduate students. It may be read easily by anyone with a working knowledge of real and complex variable theory. Topics include the finite and non-finite kernels, variation diminishing transforms, asymptotic behavior of kernels, real inversion theory, representation theory, the Weierstrass transform, and

  2. Euclidean geometry and transformations

    CERN Document Server

    Dodge, Clayton W

    1972-01-01

    This introduction to Euclidean geometry emphasizes transformations, particularly isometries and similarities. Suitable for undergraduate courses, it includes numerous examples, many with detailed answers. 1972 edition.

  3. Transform image enhancement

    Science.gov (United States)

    Aghagolzadeh, Sabzali; Ersoy, Okan K.

    1992-03-01

    Blockwise transform image enhancement techniques are discussed. Previously, transform image enhancement has usually been based on the discrete Fourier transform (DFT) applied to the whole image. Two major drawbacks with the DFT are high complexity of implementation involving complex multiplications and additions, with intermediate results being complex numbers, and the creation of severe block effects if image enhancement is done blockwise. In addition, the quality of enhancement is not very satisfactory. It is shown that the best transforms for transform image coding, namely, the scrambled real discrete Fourier transform, the discrete cosine transform, and the discrete cosine-III transform, are also the best for image enhancement. Three techniques of enhancement discussed in detail are alpha- rooting, modified unsharp masking, and filtering motivated by the human visual system response (HVS). With proper modifications, it is observed that unsharp masking and HVS- motivated filtering without nonlinearities are basically equivalent. Block effects are completely removed by using an overlap-save technique in addition to the best transform.

  4. Construction of transformation system in Penicillium purpurogenum.

    Science.gov (United States)

    Kojima, Ryo; Arai, Teppei; Kasumi, Takafumi; Ogihara, Jun

    2015-03-01

    Penicillium purpurogenum attracts attention in the food industry and biomass degradation. We expressed green fluorescent protein (GFP) with pBPE, a novel vector, and constructed a transformation system for P. purpurogenum. The accumulation of GFP was confirmed by fluorescence microscopy. In future, this system may prove useful for the genetic modification of P. purpurogenum. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  5. Genetic engineering of Geobacillus spp.

    Science.gov (United States)

    Kananavičiūtė, Rūta; Čitavičius, Donaldas

    2015-04-01

    Members of the genus Geobacillus are thermophiles that are of great biotechnological importance, since they are sources of many thermostable enzymes. Because of their metabolic versatility, geobacilli can be used as whole-cell catalysts in processes such as bioconversion and bioremediation. The effective employment of Geobacillus spp. requires the development of reliable methods for genetic engineering of these bacteria. Currently, genetic manipulation tools and protocols are under rapid development. However, there are several convenient cloning vectors, some of which replicate autonomously, while others are suitable for the genetic modification of chromosomal genes. Gene expression systems are also intensively studied. Combining these tools together with proper techniques for DNA transfer, some Geobacillus strains were shown to be valuable producers of recombinant proteins and industrially important biochemicals, such as ethanol or isobutanol. This review encompasses the progress made in the genetic engineering of Geobacillus spp. and surveys the vectors and transformation methods that are available for this genus.

  6. Agrobacterium-mediated transformation of Fusarium proliferatum.

    Science.gov (United States)

    Bernardi-Wenzel, J; Quecine, M C; Azevedo, J L; Pamphile, J A

    2016-06-03

    Fusarium proliferatum is an important pathogen that is associated with plant diseases and primarily affects aerial plant parts by producing different mycotoxins, which are toxic to humans and animals. Within the last decade, this fungus has also been described as one of the causes of red root rot or sudden death syndrome in soybean, which causes extensive damage to this crop. This study describes the Agrobacterium tumefaciens-mediated transformation of F. proliferatum as a tool for the disruption of pathogenicity genes. The genetic transformation was performed using two binary vectors (pCAMDsRed and pFAT-GFP) containing the hph (hygromycin B resistance) gene as a selection marker and red and green fluorescence, respectively. The presence of acetosyringone and the use of filter paper or nitrocellulose membrane were evaluated for their effect on the transformation efficiency. A mean processing rate of 94% was obtained with 96 h of co-cultivation only in the presence of acetosyringone and the use of filter paper or nitrocellulose membrane did not affect the transformation process. Hygromycin B resistance and the presence of the hph gene were confirmed by PCR, and fluorescence due to the expression of GFP and DsRed protein was monitored in the transformants. A high rate of mitotic stability (95%) was observed. The efficiency of Agrobacterium-mediated transformation of F. proliferatum allows the technique to be used for random insertional mutagenesis studies and to analyze fungal genes involved in the infection process.

  7. Genetic Breakthrough

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A new calf breeding technique shows promise for treating malignant tumors Chinese scientists have successfully bred a genetically altered cow capable of producing cancer-curing proteins for human beings.

  8. Transformation of technical infrastructure

    DEFF Research Database (Denmark)

    Nielsen, Susanne Balslev

    1998-01-01

    article about the need of new planning forums in order to initiate transformations with in management of large technical systems for energy, waste and water supply.......article about the need of new planning forums in order to initiate transformations with in management of large technical systems for energy, waste and water supply....

  9. Transformation optics and metamaterials

    Science.gov (United States)

    Chen, Huanyang; Chan, C. T.; Sheng, Ping

    2010-05-01

    Underpinned by the advent of metamaterials, transformation optics offers great versatility for controlling electromagnetic waves to create materials with specially designed properties. Here we review the potential of transformation optics to create functionalities in which the optical properties can be designed almost at will. This approach can be used to engineer various optical illusion effects, such as the invisibility cloak.

  10. Integral transformational coaching

    NARCIS (Netherlands)

    Keizer, W.A.J.; Nandram, S.S.

    2009-01-01

    In Chap. 12, Keizer and Nandram present the concept of Integral Transformational Coaching based on the concept of Flow and its effects on work performance. Integral Transformational Coaching is a method that prevents and cures unhealthy stress and burnout. They draw on some tried and tested spiritua

  11. Fieldwork in Transforming Societies

    DEFF Research Database (Denmark)

    Clark, Ed; Michailova, Snejina

    The contributors to this text discuss the personal and professional challenges of conducting fieldwork in the difficult, sometimes threatening contexts of the transforming societies of post-socialist Europe and China.......The contributors to this text discuss the personal and professional challenges of conducting fieldwork in the difficult, sometimes threatening contexts of the transforming societies of post-socialist Europe and China....

  12. Disc piezoelectric ceramic transformers.

    Science.gov (United States)

    Erhart, Jirií; Půlpán, Petr; Doleček, Roman; Psota, Pavel; Lédl, Vít

    2013-08-01

    In this contribution, we present our study on disc-shaped and homogeneously poled piezoelectric ceramic transformers working in planar-extensional vibration modes. Transformers are designed with electrodes divided into wedge, axisymmetrical ring-dot, moonie, smile, or yin-yang segments. Transformation ratio, efficiency, and input and output impedances were measured for low-power signals. Transformer efficiency and transformation ratio were measured as a function of frequency and impedance load in the secondary circuit. Optimum impedance for the maximum efficiency has been found. Maximum efficiency and no-load transformation ratio can reach almost 100% and 52 for the fundamental resonance of ring-dot transformers and 98% and 67 for the second resonance of 2-segment wedge transformers. Maximum efficiency was reached at optimum impedance, which is in the range from 500 Ω to 10 kΩ, depending on the electrode pattern and size. Fundamental vibration mode and its overtones were further studied using frequency-modulated digital holographic interferometry and by the finite element method. Complementary information has been obtained by the infrared camera visualization of surface temperature profiles at higher driving power.

  13. Integral transformational coaching

    NARCIS (Netherlands)

    Keizer, W.A.J.; Nandram, S.S.

    2009-01-01

    In Chap. 12, Keizer and Nandram present the concept of Integral Transformational Coaching based on the concept of Flow and its effects on work performance. Integral Transformational Coaching is a method that prevents and cures unhealthy stress and burnout. They draw on some tried and tested

  14. Flames of Transformation

    DEFF Research Database (Denmark)

    Sørensen, Tim Flohr; Bille, Mikkel

    2008-01-01

    This paper explores the transformative power of fire, its fundamental ability to change material worlds and affect our experience of its materiality. The paper examines material transformations related to death as a means of illustrating the powerful property of fire as a materially destructive y...

  15. Adaptive Wavelet Transforms

    Energy Technology Data Exchange (ETDEWEB)

    Szu, H.; Hsu, C. [Univ. of Southwestern Louisiana, Lafayette, LA (United States)

    1996-12-31

    Human sensors systems (HSS) may be approximately described as an adaptive or self-learning version of the Wavelet Transforms (WT) that are capable to learn from several input-output associative pairs of suitable transform mother wavelets. Such an Adaptive WT (AWT) is a redundant combination of mother wavelets to either represent or classify inputs.

  16. On an integral transform

    Directory of Open Access Journals (Sweden)

    D. Naylor

    1986-01-01

    Full Text Available This paper establishes properties of a convolution type integral transform whose kernel is a Macdonald type Bessel function of zero order. An inversion formula is developed and the transform is applied to obtain the solution of some related integral equations.

  17. Direct current transformer

    Science.gov (United States)

    Khanna, S. M.; Urban, E. W. (Inventor)

    1979-01-01

    A direct current transformer in which the primary consists of an elongated strip of superconductive material, across the ends of which is direct current potential is described. Parallel and closely spaced to the primary is positioned a transformer secondary consisting of a thin strip of magnetoresistive material.

  18. Incremental Distance Transforms (IDT)

    NARCIS (Netherlands)

    Schouten, Theo E.; van den Broek, Egon; Erçil, A.; Çetin, M.; Boyer, K.; Lee, S.-W.

    2010-01-01

    A new generic scheme for incremental implementations of distance transforms (DT) is presented: Incremental Distance Transforms (IDT). This scheme is applied on the cityblock, Chamfer, and three recent exact Euclidean DT (E2DT). A benchmark shows that for all five DT, the incremental implementation r

  19. Deployment & Market Transformation (Brochure)

    Energy Technology Data Exchange (ETDEWEB)

    2012-04-01

    NREL's deployment and market transformation (D and MT) activities encompass the laboratory's full range of technologies, which span the energy efficiency and renewable energy spectrum. NREL staff educates partners on how they can advance sustainable energy applications and also provides clients with best practices for reducing barriers to innovation and market transformation.

  20. Transformative Learning and Identity

    Science.gov (United States)

    Illeris, Knud

    2014-01-01

    Transformative learning has usually been defined as transformations of meaning perspectives, frames of reference, and habits of mind--as proposed initially by Jack Mezirow. However, several authors have found this definition too narrow and too cognitively oriented, and Mezirow has later emphasized that emotional and social conditions are also…

  1. Transformative environmental governance

    Science.gov (United States)

    Transformative governance is an approach to environmental governance that has the capacity to respond to, manage, and trigger regime shifts in coupled social-ecological systems (SESs) at multiple scales. The goal of transformative governance is to actively shift degraded SESs to ...

  2. The twisted Mellin transform

    OpenAIRE

    Wang, Zuoqin

    2007-01-01

    The "twisted Mellin transform" is a slightly modified version of the usual classical Mellin transform on $L^2(\\mathbb R)$. In this short note we investigate some of its basic properties. From the point of views of combinatorics one of its most important interesting properties is that it intertwines the differential operator, $df/dx$, with its finite difference analogue, $\

  3. Mitochondrial genetics

    OpenAIRE

    Chinnery, Patrick Francis; Hudson, Gavin

    2013-01-01

    Introduction In the last 10 years the field of mitochondrial genetics has widened, shifting the focus from rare sporadic, metabolic disease to the effects of mitochondrial DNA (mtDNA) variation in a growing spectrum of human disease. The aim of this review is to guide the reader through some key concepts regarding mitochondria before introducing both classic and emerging mitochondrial disorders. Sources of data In this article, a review of the current mitochondrial genetics literature was con...

  4. Matricial R-transform

    CERN Document Server

    Lenczewski, Romuald

    2011-01-01

    We study the addditon problem for strongly matricially free random variables which generalize free random variables. Using operators of Toeplitz type, we derive a linearization formula for the `matricial R-transform' related to the associated convolution. It is a linear combination of Voiculescu's R-transforms in free probability with coefficients given by internal units of the considered array of subalgebras. This allows us to view this formula as the `matricial linearization property' of the R-transform. Since strong matricial freeness unifies the main types of noncommutative independence, the matricial R-transform plays the role of a unified noncommutative analog of the logarithm of the Fourier transform for free, boolean, monotone, orthogonal, s-free and c-free independence.

  5. The transformativity approach

    DEFF Research Database (Denmark)

    Holm, Isak Winkel; Lauta, Kristian Cedervall

    2017-01-01

    During the last five to ten years, a considerable body of research has begun to explore how disasters, real and imagined, trigger social transformations. Even if the contributions to this this research stems from a multitude of academic disciplines, we argue in the article, they constitute...... an identifiable and promising approach for future disaster research. We suggest naming it the transformativity approach. Whereas the vulnerability approach explores the social causation of disasters, the transformativity approach reverses the direction of the gaze and investigates the social transformation...... brought about by disasters. Put simply, the concept of vulnerability is about the upstream causes of disaster and the concept of transformativity about the downstream effects. By discussing three recent contributions (by the historian Greg Bankoff, the legal sociologist Michelle Dauber...

  6. Transformation of Digital Ecosystems

    DEFF Research Database (Denmark)

    Henningsson, Stefan; Hedman, Jonas

    2014-01-01

    In digital ecosystems, the fusion relation between business and technology means that the decision of technical compatibility of the offering is also the decision of how to position the firm relative to the coopetive relations that characterize business ecosystems. In this article we develop...... the Digital Ecosystem Technology Transformation (DETT) framework for explaining technology-based transformation of digital ecosystems by integrating theories of business and technology ecosystems. The framework depicts ecosystem transformation as distributed and emergent from micro-, meso-, and macro- level...... coopetition. The DETT framework consists an alternative to the existing explanations of digital ecosystem transformation as the rational management of one central actor balancing ecosystem tensions. We illustrate the use of the framework by a case study of transformation in the digital payment ecosystem...

  7. 水稻转座子Pong特异性RNAi载体TPAS的构建和农杆菌介导的遗传转化条件的优化%Specific RNAi vector construction of transposon Pong in rice and Agrobacterium tumefaciens-mediated genetic transformation optimization

    Institute of Scientific and Technical Information of China (English)

    高立宏; 张春玉; 柴娜; 路云侠; 刘宝; 刘立侠

    2011-01-01

    为研究水稻转座子Pong的功能,构建了Pong的特异RNAi表达载体TPAS,并利用农杆菌介导法将这个载体转化到水稻的成熟胚愈伤组织,获得了PCR阳性的再生植株.同时通过对农杆菌侵染途径、侵染浓度和侵染时间等的研究,建立并优化了水稻的遗传转化程序,结果表明:在农杆菌侵染成熟胚愈伤组织的时间为3 min,共培养2~4 d,预培养7d,G418质量浓度为150~200 mg/L条件下筛选7d,转化率可达到30%,为最佳的遗传转化参数.研究结果为今后水稻转座子Pong功能的进一步研究提供了一定的基础和参考.%To study the function of rice transposon Pong, the research constructed the specific RNAi expressed vector TPAS of transposon Pong in rice,and transferred the vectors into the mature embryo callus of the rice cultivar Matsumae,and then obtained the regeneration of the positive PCR results by Agrobacterium tumefaciens-mediated method. To establish and optimize genetic transformation process,the different infected ways,concentrations and time of Agrobacterium were chosen. The result showed that: the infection time of Agrobacterium to mature embryo callus is 3 min,the time of coculture is 2~4 d,the pre-culture period is 7 d,all the dates were the best genetic transformation parameters, making the basis for the function of rice Tn Pong.

  8. Agrobacterium-mediated transformation of maize (Zea mays) immature embryos.

    Science.gov (United States)

    Lee, Hyeyoung; Zhang, Zhanyuan J

    2014-01-01

    Agrobacterium tumefaciens-mediated transformation is one of the most efficient and simple gene delivery systems for genetic improvement and biology studies in maize. This system has become more widely used by both public and private laboratories. However, transformation efficiencies vary greatly from laboratory to laboratory for the same genotype. Here, we illustrate our advanced Agrobacterium-mediated transformation method in Hi-II maize using simple binary vectors. The protocol utilizes immature embryos as starting explants and the bar gene as a selectable marker coupled with bialaphos as a selective agent. The protocol offers efficient transformation results with high reproducibility, provided that some experimental conditions are well controlled. This transformation method, with minor modifications, can be also employed to transform certain maize inbreds.

  9. Metabolomics of Genetically Modified Crops

    Science.gov (United States)

    Simó, Carolina; Ibáñez, Clara; Valdés, Alberto; Cifuentes, Alejandro; García-Cañas, Virginia

    2014-01-01

    Metabolomic-based approaches are increasingly applied to analyse genetically modified organisms (GMOs) making it possible to obtain broader and deeper information on the composition of GMOs compared to that obtained from traditional analytical approaches. The combination in metabolomics of advanced analytical methods and bioinformatics tools provides wide chemical compositional data that contributes to corroborate (or not) the substantial equivalence and occurrence of unintended changes resulting from genetic transformation. This review provides insight into recent progress in metabolomics studies on transgenic crops focusing mainly in papers published in the last decade. PMID:25334064

  10. Metabolomics of Genetically Modified Crops

    Directory of Open Access Journals (Sweden)

    Carolina Simó

    2014-10-01

    Full Text Available Metabolomic-based approaches are increasingly applied to analyse genetically modified organisms (GMOs making it possible to obtain broader and deeper information on the composition of GMOs compared to that obtained from traditional analytical approaches. The combination in metabolomics of advanced analytical methods and bioinformatics tools provides wide chemical compositional data that contributes to corroborate (or not the substantial equivalence and occurrence of unintended changes resulting from genetic transformation. This review provides insight into recent progress in metabolomics studies on transgenic crops focusing mainly in papers published in the last decade.

  11. Metabolomics of genetically modified crops.

    Science.gov (United States)

    Simó, Carolina; Ibáñez, Clara; Valdés, Alberto; Cifuentes, Alejandro; García-Cañas, Virginia

    2014-10-20

    Metabolomic-based approaches are increasingly applied to analyse genetically modified organisms (GMOs) making it possible to obtain broader and deeper information on the composition of GMOs compared to that obtained from traditional analytical approaches. The combination in metabolomics of advanced analytical methods and bioinformatics tools provides wide chemical compositional data that contributes to corroborate (or not) the substantial equivalence and occurrence of unintended changes resulting from genetic transformation. This review provides insight into recent progress in metabolomics studies on transgenic crops focusing mainly in papers published in the last decade.

  12. 用于虫生真菌遗传转化的苯菌灵抗性菌株筛选%Screening of Benomyl Resistant Strains for Genetic Transformation of Entomogenous Fungi

    Institute of Scientific and Technical Information of China (English)

    赵津津; 谢明; 张艳军

    2012-01-01

    发展虫生真菌的抗药性是协调生防与化防,稳定真菌杀虫剂应用效果的一个重要策略。本研究从代表性地区采集植物病原样本,分离得到35株菌株,鉴定为3属5种,即灰葡萄孢霉Botrytis cinerea、扩展青霉Penicillium expansum、指状青霉Penicillium digitatum、长柄链格孢菌Alternaria longipes和茄链格孢菌Alternaria solani。应用菌落直径法测定其对苯菌灵的抗性,结果表明:供试的11株灰葡萄孢霉菌株中,最高EC50为936.2670μg·mL^-1;12株青霉属菌株中,最高EC50为0.0384μg·mL^-1;12株链格孢菌株中,最高EC50为703.5557μg·mL^-1。其中灰霉和链格孢属菌株中有4株EC50大于600μg·mL^-1的高抗菌株,即灰葡萄孢霉的菌株SD2和SD1以及茄链格孢的菌株CC2-2和LF1-2,这4株菌株可作为遗传转化抗性基因克隆的基础材料。%One strategy to stabilize the performance of fungal pesticides in case of chemical pesticides,is to develop the resistance of the entomopathogenic fungi to fungicides.Thirty-five phytopathogenic fungal isolates were collected from different areas of China.The isolates were identified as 5 species of 3 genus,namely Botrytis cinerea,Penicillium expansum,Penicillium digitatum,Alternaria longipes and Alternaria solani,respectively.The benomyl resistance of the isolates was determined by the colony radial growth inhibition method.The maximum EC50 values of Botrytis,Penicillium and Alternria were 936.2670,0.0384 and 703.5557 μg·mL 1,respectively.Two isolates(SD1 and SD2) of B.cinerea and two isolates(CC2-2 and LF1-2) of A.solani were screened,with EC50 values of over 600 μg·mL 1.These isolates were considered as the potential materials to clone the resistant genes for the transformation of entomopahtogenic fungi.

  13. Highly efficient transformation system for Malassezia furfur and Malassezia pachydermatis using Agrobacterium tumefaciens-mediated transformation.

    Science.gov (United States)

    Celis, A M; Vos, A M; Triana, S; Medina, C A; Escobar, N; Restrepo, S; Wösten, H A B; de Cock, H

    2017-03-01

    Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M. furfur and M. pachydermatis by combining the transformation protocols of Agaricus bisporus and Cryptococcus neoformans. Optimal temperature and co-cultivation time for transformation were 5 and 7days at 19°C and 24°C, respectively. Transformation efficiency was 0.75-1.5% for M. furfur and 0.6-7.5% for M. pachydermatis. Integration of the hpt resistance cassette and gfp was verified using PCR and fluorescence microscopy, respectively. The T-DNA was mitotically stable in approximately 80% of the transformants after 10 times sub-culturing in the absence of hygromycin. Improving transformation protocols contribute to study the biology and pathophysiology of Malassezia.

  14. Challenges and advances in genetically improving trees for the plantation forestry sector

    CSIR Research Space (South Africa)

    Verryn, SD

    2010-08-30

    Full Text Available This presentation outlines the South African plantation forestry sector and its contributions and improvement in productivity, acquiring genetic diversity, challenges and advances in genetically improving trees as well as transforming the value...

  15. Fractional Radon Transform and Transform of Wigner Operator

    Institute of Scientific and Technical Information of China (English)

    FAN Hong-Yi; CHEN Jun-Hua

    2003-01-01

    Based on the Radon transform and fractional Fourier transform we introduce the fractional Radon trans-formation (FRT). We identify the transform kernel for FRT. The FRT of Wigner operator is derived, which naturallyreduces to the projector of eigenvector of the rotated quadrature in the usual Radon transform case.

  16. Stable nuclear transformation of Gonium pectorale

    Directory of Open Access Journals (Sweden)

    Hallmann Armin

    2009-07-01

    Full Text Available Abstract Background Green algae of the family Volvocaceae are a model lineage for studying the molecular evolution of multicellularity and cellular differentiation. The volvocine alga Gonium is intermediate in organizational complexity between its unicellular relative, Chlamydomonas, and its multicellular relatives with differentiated cell types, such as Volvox. Gonium pectorale consists of ~16 biflagellate cells arranged in a flat plate. The detailed molecular analysis of any species necessitates its accessibility to genetic manipulation, but, in volvocine algae, transformation procedures have so far only been established for Chlamydomonas reinhardtii and Volvox carteri. Results Stable nuclear transformation of G. pectorale was achieved using a heterologous dominant antibiotic resistance gene, the aminoglycoside 3'-phosphotransferase VIII gene (aphVIII of Streptomyces rimosus, as a selectable marker. Heterologous 3'- and 5'-untranslated flanking sequences, including promoters, were from Chlamydomonas reinhardtii or from Volvox carteri. After particle gun bombardment of wild type Gonium cells with plasmid-coated gold particles, transformants were recovered. The transformants were able to grow in the presence of the antibiotic paromomycin and produced a detectable level of the AphVIII protein. The plasmids integrated into the genome, and stable integration was verified after propagation for over 1400 colony generations. Co-transformants were recovered with a frequency of ~30–50% when cells were co-bombarded with aphVIII-based selectable marker plasmids along with unselectable plasmids containing heterologous genes. The transcription of the co-transformed, unselectable genes was confirmed. After heterologous expression of the luciferase gene from the marine copepod Gaussia princeps, which was previously engineered to match the codon usage in C. reinhardtii, Gonium transformants show luciferase activity through light emission in bioluminescence

  17. A Simplified Seed Transformation Method for Obtaining Transgenic Brassica napus Plants

    Institute of Scientific and Technical Information of China (English)

    SONG Li; ZHAO De-gang; WU Yong-jun; TIAN Xiao-e

    2009-01-01

    We report here a seed transformation of sonication-assisted,no-tissue culture to rapidly produce transgenic Brassica napus plants.This method comprises the steps of treating seeds by ultrasonic wave,inoculating Agrobacterium tumefaciens with a recombinant ChlFN-a gene and germinating directly of treatment seed on wet filter papers.The obtained transformants were verified by GUS histochemical assay and nested PCR amplification.It suggests that seed transformation has a potential use in genetic transformation of rape.

  18. Cloning and Genetic Transformation of the Candidate Genes for the Bacterial Blight Resistance Gene Xa31(t)in Rice%水稻白叶枯病抗性基因Xa31(t候选基因的克隆及遗传转化

    Institute of Scientific and Technical Information of China (English)

    杨青; 王春台; 刘学群; 谭艳平

    2011-01-01

    To construct the expression vector with the candidate genes of bacteria blight disease resistance gene Xa31(t) in rice, and to make the genetic transformation, the candidate genes Xa3l(t)-1 and Xa31(t)-2 of rice bacterial blight resistance gene Xa31(t) were obtained through the method of long segments PCR with the resistant variety 'Zachanglong' (ZCL) genomic DNA as the template. Then the target genes were cloned to the T-vector with the way of restriction endonuclease reaction and enzyme ligation reaction. After screening the positive clones and sequencing, the target genes were cloned to the expression vector of pCMABIA1300. Finally, the recombinant plasmids were transformed into the callus of the susceptible varieties ' Nipponbare' and 'Taibei309' respectively by agrobacterium-mediated approach and a lot of transformed rice seedlings were obtained. These work laid the foundation for cloning the bacterial blight resistance gene Xa31(t) and studied its function.%为了构建水稻白叶枯病抗性基因Xa31(t)候选基因的植物表达载体并对其进行遗传转化,以水稻白叶枯病抗性品种‘扎昌龙’(ZCL)的基因组DNA为模板,通过长片段PCR扩增得到水稻白叶枯病抗性基因Xa31(t)候选基因Xa31(t)-1和Xa31(t)-2的序列,采用酶切、连接的方法先将目的基因克隆至T载体,筛选正确的质粒后,再克隆至pCMABIA1300表达载体.以农杆菌介导法将构建好的重组质粒转入‘日本睛’和‘台北309’的愈伤后诱导成苗,获得了大量转基因植株,这些工作为克隆白叶枯病抗性基因并研究其功能奠定了基础.

  19. Transformation of phaG and phaC Genes into Tobacco Chloroplast Genome and Genetic Analysis%phaG和phaC基因在烟草叶绿体中的转化及其遗传分析

    Institute of Scientific and Technical Information of China (English)

    王玉华; 吴忠义; 张秀海; 王永勤; 黄丛林; 贾敬芬

    2009-01-01

    present, novel efforts are focused on using the transgenic plants as bioreactors to produce PHAs. Both 3-hydroxyacyl-CoA-ACP-transferase and type Ⅱ PHA synthase are the key enzymes for mcl-PHAs biosynthesis. The gene phaG encoding 3-hydroxyacyl-CoA-ACP-transferase was placed under the control of psbA-pro and psbA-ter of rice to construct phaG expression cassette, and the gene phaC encoding type Ⅱ PHA synthase was placed under the control of prm and rbcL-ter of rice to construct phaC expression cassette, which were ligated with the screening marker gene aadA expression cassette prm-aadA-TpsbA-ter. These recombined fragments were cloned between the plastid rbcL and accD genes of tobacco for targeting to the large single copy region of chloroplast genome. Chloroplast expression vector of pTGC was constructed and then transformed into tobacco chloroplast genome through particle bombardment. Six trans-plastomic tobacco plants were obtained by spectinomycin screening. PCR and Southern blot analysis confirmed integration of phaG andphaC genes into chloroplast genome of T_0 and T_1 transgenic plants, and T_1 transgenic plants exhibited homogenization. The expression of phaC and phaG at transcription level was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Recombinant transgenes in the tobacco chloroplast genome were maternally inherited and were not transmitted via pollen when out-crossed with untransformed female plants.

  20. Advancing Crop Transformation in the Era of Genome Editing.

    Science.gov (United States)

    Altpeter, Fredy; Springer, Nathan M; Bartley, Laura E; Blechl, Ann E; Brutnell, Thomas P; Citovsky, Vitaly; Conrad, Liza J; Gelvin, Stanton B; Jackson, David P; Kausch, Albert P; Lemaux, Peggy G; Medford, June I; Orozco-Cárdenas, Martha L; Tricoli, David M; Van Eck, Joyce; Voytas, Daniel F; Walbot, Virginia; Wang, Kan; Zhang, Zhanyuan J; Stewart, C Neal

    2016-07-01

    Plant transformation has enabled fundamental insights into plant biology and revolutionized commercial agriculture. Unfortunately, for most crops, transformation and regeneration remain arduous even after more than 30 years of technological advances. Genome editing provides novel opportunities to enhance crop productivity but relies on genetic transformation and plant regeneration, which are bottlenecks in the process. Here, we review the state of plant transformation and point to innovations needed to enable genome editing in crops. Plant tissue culture methods need optimization and simplification for efficiency and minimization of time in culture. Currently, specialized facilities exist for crop transformation. Single-cell and robotic techniques should be developed for high-throughput genomic screens. Plant genes involved in developmental reprogramming, wound response, and/or homologous recombination should be used to boost the recovery of transformed plants. Engineering universal Agrobacterium tumefaciens strains and recruiting other microbes, such as Ensifer or Rhizobium, could facilitate delivery of DNA and proteins into plant cells. Synthetic biology should be employed for de novo design of transformation systems. Genome editing is a potential game-changer in crop genetics when plant transformation systems are optimized.

  1. Genetic Manipulations in Dermatophytes.

    Science.gov (United States)

    Alshahni, Mohamed Mahdi; Yamada, Tsuyoshi

    2017-02-01

    Dermatophytes are a group of closely related fungi that nourish on keratinized materials for their survival. They infect stratum corneum, nails, and hair of human and animals, accounting the largest portion of fungi causing superficial mycoses. Huge populations are suffering from dermatophytoses, though the biology of these fungi is largely unknown yet. Reasons are partially attributed to the poor amenability of dermatophytes to genetic manipulation. However, advancements in this field over the last decade made it possible to conduct genetic studies to satisfying extents. These included genetic transformation methods, indispensable molecular tools, i.e., dominant selectable markers, inducible promoter, and marker recycling system, along with improving homologous recombination frequency and gene silencing. Furthermore, annotated genome sequences of several dermatophytic species have recently been available, ensuring an optimal recruitment of the molecular tools to expand our knowledge on these fungi. In conclusion, the establishment of basic molecular tools and the availability of genomic data will open a new era that might change our understanding on the biology and pathogenicity of this fungal group.

  2. A Selective CPS Transformation

    DEFF Research Database (Denmark)

    Nielsen, Lasse Riechstein

    2001-01-01

    The CPS transformation makes all functions continuation-passing, uniformly. Not all functions, however, need continuations: they only do if their evaluation includes computational effects. In this paper we focus on control operations, in particular "call with current continuation" and "throw". We...... characterize this involvement as a control effect and we present a selective CPS transformation that makes functions and expressions continuation-passing if they have a control effect, and that leaves the rest of the program in direct style. We formalize this selective CPS transformation with an operational...

  3. Developing Global Transformational Leaders

    DEFF Research Database (Denmark)

    Ramsey, Jase R.; Rutti, Raina M.; Lorenz, Melanie P.

    2016-01-01

    Despite significant increases in training and development of global managers, little is known about the precursors of transformational leadership in Multilatinas. While prior cross-cultural literature suggests that being an autocratic leader is ideal in Multilatinas, using transformational...... of transformational leadership because they are better able to understand the differences of other cultures, and appropriately adjust their behavior....... leadership theory, we argue that global leaders of Multilatinas embrace a more humanistic approach to leadership because of the importance of relationships between leaders and their followers. Additionally, we argue that global leaders with high levels of cultural intelligence will have high levels...

  4. Laplace transforms essentials

    CERN Document Server

    Shafii-Mousavi, Morteza

    2012-01-01

    REA's Essentials provide quick and easy access to critical information in a variety of different fields, ranging from the most basic to the most advanced. As its name implies, these concise, comprehensive study guides summarize the essentials of the field covered. Essentials are helpful when preparing for exams, doing homework and will remain a lasting reference source for students, teachers, and professionals. Laplace Transforms includes the Laplace transform, the inverse Laplace transform, special functions and properties, applications to ordinary linear differential equations, Fourier tr

  5. A Selective CPS Transformation

    DEFF Research Database (Denmark)

    Nielsen, Lasse Riechstein

    2001-01-01

    The CPS transformation makes all functions continuation-passing, uniformly. Not all functions, however, need continuations: they only do if their evaluation includes computational effects. In this paper we focus on control operations, in particular "call with current continuation" and "throw". We...... characterize this involvement as a control effect and we present a selective CPS transformation that makes functions and expressions continuation-passing if they have a control effect, and that leaves the rest of the program in direct style. We formalize this selective CPS transformation with an operational...

  6. Solid phase transformations II

    CERN Document Server

    Čermák, J

    2009-01-01

    This topical volume includes ten invited papers that cover selected areas of the field of solid phase transformations. The first two contributions represent a burgeoning branch; that of the computer simulation of physical phenomena. The following three articles deal with the thermodynamics of phase transformations as a basic theory for describing the phenomenology of phase changes in matter. The next paper describes the interconnections between structural stability and the electronic structure of phases. Two further articles are devoted to displacive transformations; a field where there are ma

  7. Transformers and motors

    CERN Document Server

    Shultz, George

    1991-01-01

    Transformers and Motors is an in-depth technical reference which was originally written for the National Joint Apprenticeship Training Committee to train apprentice and journeymen electricians. This book provides detailed information for equipment installation and covers equipment maintenance and repair. The book also includes troubleshooting and replacement guidelines, and it contains a minimum of theory and math.In this easy-to-understand, practical sourcebook, you'll discover:* Explanations of the fundamental concepts of transformers and motors* Transformer connections and d

  8. Genetic GIScience

    DEFF Research Database (Denmark)

    Jacquez, Geoffrey; Sabel, Clive E; Shi, Chen

    2015-01-01

    The exposome, defined as the totality of an individual's exposures over the life course, is a seminal concept in the environmental health sciences. Although inherently geographic, the exposome as yet is unfamiliar to many geographers. This article proposes a place-based synthesis, genetic...... geographic information science (genetic GIScience), that is founded on the exposome, genome+, and behavome. It provides an improved understanding of human health in relation to biology (the genome+), environmental exposures (the exposome), and their social, societal, and behavioral determinants (the behavome......). Genetic GIScience poses three key needs: first, a mathematical foundation for emergent theory; second, process-based models that bridge biological and geographic scales; third, biologically plausible estimates of space?time disease lags. Compartmental models are a possible solution; this article develops...

  9. Agrobacterium-mediated transformation of ornamental species: A review

    Directory of Open Access Journals (Sweden)

    Milošević Snežana

    2015-01-01

    Full Text Available Integration of desirable traits into commercial ornamentals using genetic engineering techniques is a powerful tool in contemporary biotechnology. However, these techniques have had a limited impact in the domain of ornamental horticulture, particularly floriculture. Modifications of the color, architecture or fragrance of the flowers as well as an improvement of the plant tolerance/resistance against abiotic and biotic stresses using plant transformation techniques, is still in its infancy. This review focuses on the application of Agrobacterium-mediated transformation, a major plant genetic engineering approach to ornamental plant breeding and the impact it has had to date. [Projekat Ministarstva nauke Republike Srbije, br. TR 31019

  10. Reubicación del parque de transformadores de los sistemas de distribución de Bogotá D.C. mediante algoritmos genéticos Relocation of electric transformers lot in Bogotá distribution systems using genetic algorithms

    Directory of Open Access Journals (Sweden)

    Johnn Alejandro Quintero Salazar

    2012-08-01

    Full Text Available Este documento presenta una metodología basada en algoritmos genéticos que permite adelantar el reordenamiento del parque de transformadores existente en el nivel de tensión I de los sistemas de distribución de Bogotá D.C., Colombia, con el fin de maximizar el reconocimiento de activos que el ente regulador CREG (Comisión Reguladora de Energía y Gas realiza a los distintos operadores de red, según lo establecido en la resolución 097 de 2008. Para la aplicación del algoritmo se obtuvieron mediciones de potencia activa máxima para cada hora del año 2009 en un conjunto de 94 transformadores de diferentes capacidades, escogidos de forma aleatoria, instalados en el sistema de distribución de CODENSA S.A. ESP, empresa encargada de prestar el servicio de energía eléctrica en la ciudad de Bogotá D.C. Con esta información se construyeron curvas de carga diarias representativas y se elaboró una base de datos que contiene los costos operativos del movimiento de los equipos y de las tarifas, a partir de los cuales fue posible modelar la función objetivo y las restricciones del problema, obteniéndose un elevado número de combinaciones posibles (alrededor de 1*10(134 debido al gran número de nodos y de transformadores presentes en el sistema de distribución. La búsqueda convencional de una solución en la anterior situación implicaría el empleo de tiempos prohibitivos, por lo cual se implementó un algoritmo genético clásico, obteniéndose de esta manera una solución óptima que ofrece una ganancia económica en el primer año, asociada al incremento en el cargo por uso, de $ 253.446.362,47 (COP, ganancias que podrían verse incrementadas considerablemente al ejecutar el algoritmo en parques de transformadores más grandes.This paper presents a methodology based on genetic algorithms that allows the reordering of the existing park transformers voltage level I of the distribution systems of Bogotá city, Colombia, in order to

  11. Influence of antibiotics on embryogenic tissue and Agrobacterium tumefaciens suppression in soybean genetic transformation Influência de antibióticos sobre o tecido embriogênico e a supressão de Agrobacterium tumefaciens na transformação genética de soja

    Directory of Open Access Journals (Sweden)

    Beatriz Wiebke

    2006-01-01

    Full Text Available The influence of different antibiotic treatments in soybean genetic transformation was evaluated. First, an assay was performed to verify how different antibiotic treatments affect soybean embryogenic tissues. The effect of carbenicillin at 500 mg L-1 was genotype-dependent. This antibiotic did not affect embryo survival of cv. IAS5, but a three-fold increase of embryo proliferation was observed for cv. Bragg, when compared to the control. On the other hand, cefotaxime at 350 and 500 mg L-1 caused death of embryogenic tissues of both cultivars. Finally, the association of cefotaxime (250 mg L-1 + vancomycin (250 mg L-1 did not affect negatively the somatic embryos of tested cultivars until 63 days of treatment. Thereafter, a second experiment was carried out to determine the efficacy of different antibiotic treatments in suppressing LBA4404 Agrobacterium tumefaciens strain in genetic transformation. On tissue culture conditions, carbenicillin at 500 and 1000 mg L-1 was not active against Agrobacterium. On the other hand, treatments with cefotaxime at 350 and 500 mg L-1, and cefotaxime + vancomycin efficiently suppressed Agrobacterium during 49 days. Data of both experiments suggested cefotaxime + vancomycin for 49-63 days as the most appropriate treatment. This is the first work reporting the effect of antibiotics on soybean tissues. By identifying an antibiotic combination that suppressed A. tumefaciens with minimal phytotoxic effects, we are able to recommend it for improvement of soybean Agrobacterium-mediated transformation procedure.Foi avaliada a influência de diferentes tratamentos com antibióticos durante a transformação genética de soja. Inicialmente, desenvolveu-se um estudo para identificar como diferentes tratamentos com antibióticos afetam o tecido embriogênico da soja. O efeito da carbenicilina a 500 mg L-1 foi genótipo-dependente. Esse antibiótico não afetou a sobrevivência dos embriões da cv. IAS5, enquanto a

  12. Matrices and transformations

    CERN Document Server

    Pettofrezzo, Anthony J

    1978-01-01

    Elementary, concrete approach: fundamentals of matrix algebra, linear transformation of the plane, application of properties of eigenvalues and eigenvectors to study of conics. Includes proofs of most theorems. Answers to odd-numbered exercises.

  13. Transforming Homeland Security [video

    OpenAIRE

    McIntyre, David; Center for Homeland Defense and Security Naval Postgraduate School

    2011-01-01

    A pioneer in homeland security, and homeland security education, David McIntyre discusses the complexities in transforming homeland security from a national program in its inception, to also include state and local agencies and other public and private parties.

  14. Transformer design tradeoffs

    Science.gov (United States)

    Mclyman, W. T.

    1977-01-01

    In space, power system transformer components are frequently the heaviest and bulkiest items in the power conversion circuit. They also have a significant effect upon the overall performance and efficiency of the system. Accordingly, the design of such transformers has an important effect on overall system weight, power-inversion efficiency, and cost. Relationships were between the parameters used by transformer designers that can be used as new tools to standardize and simplify transformer design. They can be used to optimize the design either for small size and weight or efficiency. The metric system of units, rather than the familiar English units, is used; however, material is presented to assist the reader in the transition from one system to the other.

  15. Finite BMS transformations

    Energy Technology Data Exchange (ETDEWEB)

    Barnich, Glenn [Physique Théorique et Mathématique,Université Libre de Bruxelles and International Solvay Institutes,Campus Plaine C.P. 231, B-1050 Bruxelles (Belgium); Troessaert, Cédric [Centro de Estudios Científicos (CECs),Arturo Prat 514, Valdivia (Chile)

    2016-03-24

    The action of finite BMS and Weyl transformations on the gravitational data at null infinity is worked out in three and four dimensions in the case of an arbitrary conformal factor for the boundary metric induced on Scri.

  16. On noncommutative Nahm transform

    Energy Technology Data Exchange (ETDEWEB)

    Astashkevich, A.; Schwarz, A. [California Univ., Davis, CA (United States). Dept. of Mathematics; Nekrasov, N. [Lyman Laboratory of Physics, Harvard University, Cambridge, MA (United States); Institute of Theoretical and Experimental Physics, Moscow (Russian Federation)

    2000-04-01

    Motivated by the recently observed relation between the physics of D-branes in the background of B-field and the noncommutative geometry we study the analogue of the Nahm transform for the instantons on the noncommutative torus. (orig.)

  17. Fourier transform mass spectrometry.

    Science.gov (United States)

    Scigelova, Michaela; Hornshaw, Martin; Giannakopulos, Anastassios; Makarov, Alexander

    2011-07-01

    This article provides an introduction to Fourier transform-based mass spectrometry. The key performance characteristics of Fourier transform-based mass spectrometry, mass accuracy and resolution, are presented in the view of how they impact the interpretation of measurements in proteomic applications. The theory and principles of operation of two types of mass analyzer, Fourier transform ion cyclotron resonance and Orbitrap, are described. Major benefits as well as limitations of Fourier transform-based mass spectrometry technology are discussed in the context of practical sample analysis, and illustrated with examples included as figures in this text and in the accompanying slide set. Comparisons highlighting the performance differences between the two mass analyzers are made where deemed useful in assisting the user with choosing the most appropriate technology for an application. Recent developments of these high-performing mass spectrometers are mentioned to provide a future outlook.

  18. Distributional Watson transforms

    NARCIS (Netherlands)

    Dijksma, A.; Snoo, H.S.V. de

    1974-01-01

    For all Watson transforms W in L2(R+) a triple of Hilbert space LG ⊂ L2(R+) ⊂ L'G is constructed such that W may be extended to L'G. These results allow the construction of a triple L ⊂ L2(R+) ⊂ L', where L is a Gelfand-Fréchet space. This leads to a theory of distributional Watson transforms.

  19. Power transformers quality assurance

    CERN Document Server

    Dasgupta, Indrajit

    2009-01-01

    About the Book: With the view to attain higher reliability in power system operation, the quality assurance in the field of distribution and power transformers has claimed growing attention. Besides new developments in the material technology and manufacturing processes of transformers, regular diagnostic testing and maintenance of any engineering product may be ascertained by ensuring: right selection of materials and components and their quality checks. application of correct manufacturing processes any systems engineering. the user`s awareness towards preventive maintenance. The

  20. Transformational VLSI Design

    DEFF Research Database (Denmark)

    Rasmussen, Ole Steen

    This thesis introduces a formal approach to deriving VLSI circuits by the use of correctness-preserving transformations. Both the specification and the implementation are descibed by the relation based language Ruby. In order to prove the transformation rules a proof tool called RubyZF has been...... in connection with VLSI design are defined in terms of Pure Ruby and their properties proved. The design process is illustrated by several non-trivial examples of standard VLSI problems....

  1. Process for compound transformation

    KAUST Repository

    Basset, Jean-Marie

    2016-12-29

    Embodiments of the present disclosure provide for methods of using a catalytic system to chemically transform a compound (e.g., a hydrocarbon). In an embodiment, the method does not employ grafting the catalyst prior to catalysis. In particular, embodiments of the present disclosure provide for a process of hydrocarbon (e.g., C1 to C20 hydrocarbon) metathesis (e.g., alkane, olefin, or alkyne metathesis) transformation, where the process can be conducted without employing grafting prior to catalysis.

  2. Fourier transformation for pedestrians

    CERN Document Server

    Butz, Tilman

    2006-01-01

    Meant to serve an "entertaining textbook," this book belongs to a rare genre. It is written for all students and practitioners who deal with Fourier transformation. Fourier series as well as continuous and discrete Fourier transformation are covered, and particular emphasis is placed on window functions. Many illustrations and easy-to-solve exercises make the book especially accessible, and its humorous style will add to the pleasure of learning from it.

  3. The Multidimensional Darboux Transformation

    CERN Document Server

    González-López, A; González-López, Artemio; Kamran, Niky

    1996-01-01

    A generalization of the classical one-dimensional Darboux transformation to arbitrary n-dimensional oriented Riemannian manifolds is constructed using an intrinsic formulation based on the properties of twisted Hodge Laplacians. The classical two-dimensional Moutard transformation is also generalized to non-compact oriented Riemannian manifolds of dimension n greater than one. New examples of quasi-exactly solvable multidimensional matrix Schrödinger operators on curved manifolds are obtained by applying the above results.

  4. Generalization of Hasimoto's transformation

    CERN Document Server

    Molitor, Mathieu

    2012-01-01

    In this paper, we generalize the famous Hasimoto's transformation by showing that the dynamics of a closed unidimensional vortex filament embedded in a three-dimensional manifold of constant curvature gives rise under Hasimoto's transformation to the non-linear Schrodinger equation. We also give a natural interpretation of the function \\psi introduced by Hasimoto in terms of moving frames associated to a natural complex bundle over the filament.

  5. Structural Transformation in Ecuador

    OpenAIRE

    Ricardo Hausmann; Bailey Klinger

    2010-01-01

    This paper applies new techniques and metrics to analyze Ecuador's past record of and future opportunities for structural transformation. Ecuador's export dynamics and the emergence of new export activities have been the historical drivers of the country's growth, but recently Ecuador's export basket has undergone little structural transformation. The same broad sectors continue to dominate, and the overall sophistication of the export basket has actually declined in recent years. In order to...

  6. Highly efficient sorghum transformation

    OpenAIRE

    Liu, Guoquan; Godwin, Ian D.

    2012-01-01

    A highly efficient microprojectile transformation system for sorghum (Sorghum bicolor L.) has been developed by using immature embryos (IEs) of inbred line Tx430. Co-bombardment was performed with the neomycin phosphotransferase II (nptII) gene and the green fluorescent protein (gfp) gene, both under the control of the maize ubiquitin1 (ubi1) promoter. After optimization of both tissue culture media and parameters of microprojectile transformation, 25 independent transgenic events were obtain...

  7. Business process transformation

    CERN Document Server

    Grover, Varun

    2015-01-01

    Featuring contributions from prominent thinkers and researchers, this volume in the ""Advances in Management Information Systems"" series provides a rich set of conceptual, empirical, and introspective studies that epitomize fundamental knowledge in the area of Business Process Transformation. Processes are interpreted broadly to include operational and managerial processes within and between organizations, as well as those involved in knowledge generation. Transformation includes radical and incremental change, its conduct, management, and outcome. The editors and contributing authors pay clo

  8. Series Transmission Line Transformer

    Science.gov (United States)

    Buckles, Robert A.; Booth, Rex; Yen, Boris T.

    2004-06-29

    A series transmission line transformer is set forth which includes two or more of impedance matched sets of at least two transmissions lines such as shielded cables, connected in parallel at one end ans series at the other in a cascading fashion. The cables are wound about a magnetic core. The series transmission line transformer (STLT) which can provide for higher impedance ratios and bandwidths, which is scalable, and which is of simpler design and construction.

  9. Bar piezoelectric ceramic transformers.

    Science.gov (United States)

    Erhart, Jiří; Pulpan, Půlpán; Rusin, Luboš

    2013-07-01

    Bar-shaped piezoelectric ceramic transformers (PTs) working in the longitudinal vibration mode (k31 mode) were studied. Two types of the transformer were designed--one with the electrode divided into two segments of different length, and one with the electrodes divided into three symmetrical segments. Parameters of studied transformers such as efficiency, transformation ratio, and input and output impedances were measured. An analytical model was developed for PT parameter calculation for both two- and three-segment PTs. Neither type of bar PT exhibited very high efficiency (maximum 72% for three-segment PT design) at a relatively high transformation ratio (it is 4 for two-segment PT and 2 for three-segment PT at the fundamental resonance mode). The optimum resistive loads were 20 and 10 kΩ for two- and three-segment PT designs for the fundamental resonance, respectively, and about one order of magnitude smaller for the higher overtone (i.e., 2 kΩ and 500 Ω, respectively). The no-load transformation ratio was less than 27 (maximum for two-segment electrode PT design). The optimum input electrode aspect ratios (0.48 for three-segment PT and 0.63 for two-segment PT) were calculated numerically under no-load conditions.

  10. Floral Transformation of Wheat

    Science.gov (United States)

    Agarwal, Sujata; Loar, Star; Steber, Camille; Zale, Janice

    A method is described for the floral transformation of wheat using a protocol similar to the floral dip of Arabidopsis. This method does not employ tissue culture of dissected embryos, but instead pre-anthesis spikes with clipped florets at the early, mid to late uninucleate microspore stage are dipped in Agrobacterium infiltration media harboring a vector carrying anthocyanin reporters and the NPTII selectable marker. T1 seeds are examined for color changes induced in the embryo by the anthocyanin reporters. Putatively transformed seeds are germinated and the seedlings are screened for the presence of the NPTII gene based on resistance to paromomycin spray and assayed with NPTII ELISAs. Genomic DNA of putative transformants is digested and analyzed on Southern blots for copy number to determine whether the T-DNA has integrated into the nucleus and to show the number of insertions. The non-optimized transformation efficiencies range from 0.3 to 0.6% (number of transformants/number of florets dipped) but the efficiencies are higher in terms of the number of transformants produced/number of seeds set ranging from 0.9 to 10%. Research is underway to maximize seed set and optimize the protocol by testing different Agrobacterium strains, visual reporters, vectors, and surfactants.

  11. Pharmacogenetics: transforming clinical medicine.

    Science.gov (United States)

    Newman, W G

    2012-01-01

    Pharmacogenetics, the study of genetic variation relevant to drug metabolism, is a rapidly evolving area of medicine. This brief review will consider some of the recent advances where inherited genetic variants have been associated with either drug efficacy or toxicity. Examples of where pharmacogenetic testing has been adopted into clinical practice will be provided as well as a look at its likely development over the next decade. Finally, the large increase in genetic testing of tumour tissue samples to predict response to molecularly targeted treatments in cancer will be considered.

  12. Transformers analysis, design, and measurement

    CERN Document Server

    Lopez-Fernandez, Xose M; Turowski, Janusz

    2012-01-01

    This book focuses on contemporary economic, design, diagnostics, and maintenance aspects of power, instrument, and high frequency transformers, which are critical to designers for a transformer stations. The text covers such topics as shell type and superconducting transformers as well as coreless PCB and planar transformers. It emphasizes challenges and strategies in transformer design and illustrates the importance of economics in transformers management by reviewing life cycle cost design and the use of decision methods to manage risk.

  13. PLANT TISSUE CULTURE IN BIOTECHNOLOGY: RECENT ADVANCES IN TRANSFORMATION THROUGH SOMATIC EMBRYOGENESIS

    Directory of Open Access Journals (Sweden)

    V. A. Sidorov

    2012-08-01

    Full Text Available Plant genetic transformation has become an important biotechnology tool for the improvement of many crops. A solid foundation for the fast development and implementation of biotechnology in agriculture was provided by achievements in plant tissue culture. On the 30th anniversary of plant transformation, I report the advancements, recent challenges and shifts in methodology of transformation. The main focus of this paper will be on conventional and novel approaches for genetic improvements of soybean, cotton and corn. I will also highlight results on the transformation of these crops that have considerably been improved by modern biotechnology.

  14. RNA genetics

    Energy Technology Data Exchange (ETDEWEB)

    Domingo, E. (Instituto de Biologia Molecular, Facultad de Ciencias, Universidad Autonoma de Madrid, Canto Blanco, Madrid (ES)); Holland, J.J. (California Univ., San Diego, La Jolla, CA (USA). Dept. of Biology); Ahlquist, P. (Wisconsin Univ., Madison, WI (USA). Dept. of Plant Pathology)

    1988-01-01

    This book contains the proceedings on RNA genetics: RNA-directed virus replication Volume 1. Topics covered include: Replication of the poliovirus genome; Influenza viral RNA transcription and replication; and Relication of the reoviridal: Information derived from gene cloning and expression.

  15. Genetic counseling

    Science.gov (United States)

    ... MF, eds. Creasy and Resnik's Maternal-Fetal Medicine: Principles and Practice . 7th ed. Philadelphia, PA: Elsevier Saunders; 2014:chap 30. Review Date 1/25/2016 Updated by: Chad Haldeman-Englert, MD, FACMG, Fullerton Genetics Center, Asheville, NC. Review provided by VeriMed Healthcare ...

  16. Recent Advances in Genetic Engineering - A Review

    OpenAIRE

    Sobiah Rauf; Zubair Anwar; Hussain Mustatab Wahedi; Jabar Zaman Khan Khattak; Talal Jamil

    2012-01-01

    Humans have been doing genetic engineering, a technology which is transforming our world, for thousands of years on a wide range of plants, animals and micro organism and have applications in the field of medicine, research, industry and agriculture. The rapid developments in the field of genetic engineering have given a new impetus to biotechnology. This introduces the possibility of tailoring organisms in order to optimize the production of established or novel metabolites of commercial imp...

  17. Transformers: analysis, design, and measurement

    National Research Council Canada - National Science Library

    López-Fernández, Xose M; Ertan, H. Bülent; Turowski, J

    2013-01-01

    "This book focuses on contemporary economic, design, diagnostics, and maintenance aspects of power, instrument, and high frequency transformers, which are critical to designers for a transformer stations...

  18. Legendre transforms in chemical thermodynamics

    Energy Technology Data Exchange (ETDEWEB)

    Alberty, R.A. (Massachusetts Inst. of Technology, Cambridge, MA (United States). Dept. of Chemistry)

    After introductory remarks on chemical thermodynamics, independent variables, and natural variables, the paper discusses the following: Callen's nomenclature for Legendre transformed thermodynamic potentials; Transforms for chemical work (gas reactions, biochemical reactions, and ligand binding and denaturation of macromolecules); Transforms for gravitational and centrifugal work; Transforms for mechanical work (tensile stress and shear stress); Transforms for surface work; Transforms for work of electrical transport (fundamental equations, thermodynamic properties and chemical reactions, and derivation of the equation for the membrane potential); Transforms for work of electric polarization; and Transforms for work of magnetic polarization. 92 references.

  19. Agrobacterium tumefaciens-mediated transformation of CryⅠA(b) gene to Trichoderma harzianum

    Institute of Scientific and Technical Information of China (English)

    GAO Xingxi; YANG Qian

    2004-01-01

    In this study, CryⅠA(b) gene was successfully transferred into the biocontrol fungus Trichoderma harzianum with an efficiency of 60-180 transformants per 106 spores by using Agrobacterium tumefaciens-mediated trans- formation. Putative transformants were analyzed to test the presence of CryⅠA(b) gene by Southern blot. Most transformants contained a single T-DNA copy. RT-PCR analysis showed that the CryⅠA(b) gene was transcribed. Antifungal activities and insecticidal activities of the transformants were examined. There was no obvious difference in antifungal activities between the transformants and their wild strains. The modified mortalities of the transformants T1 and T2 were 69.57% and 91.30%, respectively. The tranformation system mediated by A. tumefaciens proved to be a powerful tool for the filamentous fungi transformation and functional genomic study with its high transformation frequency, simplicity of T-DNA integration, and genetic stability of transformants.

  20. From Complex Fractional Fourier Transform to Complex Fractional Radon Transform

    Institute of Scientific and Technical Information of China (English)

    FAN Hong-Yi; JIANG Nian-Quan

    2004-01-01

    We show that for n-dimensional complex fractional Fourier transform the corresponding complex fractional Radon transform can also be derived, however, it is different from the direct product of two n-dimensional real fractional Radon transforms. The complex fractional Radon transform of two-mode Wigner operator is calculated.

  1. Phase Transformations and Earthquakes

    Science.gov (United States)

    Green, H. W.

    2011-12-01

    Phase transformations have been cited as responsible for, or at least involved in, "deep" earthquakes for many decades (although the concept of "deep" has varied). In 1945, PW Bridgman laid out in detail the string of events/conditions that would have to be achieved for a solid/solid transformation to lead to a faulting instability, although he expressed pessimism that the full set of requirements would be simultaneously achieved in nature. Raleigh and Paterson (1965) demonstrated faulting during dehydration of serpentine under stress and suggested dehydration embrittlement as the cause of intermediate depth earthquakes. Griggs and Baker (1969) produced a thermal runaway model of a shear zone under constant stress, culminating in melting, and proposed such a runaway as the origin of deep earthquakes. The discovery of Plate Tectonics in the late 1960s established the conditions (subduction) under which Bridgman's requirements for earthquake runaway in a polymorphic transformation could be possible in nature and Green and Burnley (1989) found that instability during the transformation of metastable olivine to spinel. Recent seismic correlation of intermediate-depth-earthquake hypocenters with predicted conditions of dehydration of antigorite serpentine and discovery of metastable olivine in 4 subduction zones, suggests strongly that dehydration embrittlement and transformation-induced faulting are the underlying mechanisms of intermediate and deep earthquakes, respectively. The results of recent high-speed friction experiments and analysis of natural fault zones suggest that it is likely that similar processes occur commonly during many shallow earthquakes after initiation by frictional failure.

  2. Study of wavelet transform type high-current transformer

    Institute of Scientific and Technical Information of China (English)

    卢文科; 朱长纯; 刘君华; 张建军

    2002-01-01

    The wavelet transformation is applied to the high-current transformer.The high-current transformer elaborated in the paper is mainly applied to the measurement of AC/DC high-current.The principle of the transformer is the Hall direct-measurement principle.The transformer has the following three characteristics:firstly, the effect of the remnant field of the iron core on the measurement is decreased;secondly,because the temperature compensation is adopted,the transformer has good temperature charactreristic;thirdly,be-cause the wavelet transfomation technology is adopted,the transformer has the capacity of good antijanming.

  3. Nonlocal transformation optics

    CERN Document Server

    Castaldi, Giuseppe; Alu', Andrea; Engheta, Nader

    2011-01-01

    We show that the powerful framework of transformation optics may be exploited for engineering the nonlocal response of artificial electromagnetic materials. Relying on the form-invariant properties of coordinate-transformed Maxwell's equations in the spectral domain, we derive the general constitutive "blueprints" of transformation media yielding prescribed nonlocal field-manipulation effects, and provide a physically-incisive and powerful geometrical interpretation in terms of deformation of the equi-frequency contours. In order to illustrate the potentials of our approach, we present an example of application to a wave-splitting refraction scenario, which may be implemented via a simple class of artificial materials. Our results provide a systematic and versatile framework which may open intriguing venues in dispersion engineering of artificial materials.

  4. Invariants of polarization transformations.

    Science.gov (United States)

    Sadjadi, Firooz A

    2007-05-20

    The use of polarization-sensitive sensors is being explored in a variety of applications. Polarization diversity has been shown to improve the performance of the automatic target detection and recognition in a significant way. However, it also brings out the problems associated with processing and storing more data and the problem of polarization distortion during transmission. We present a technique for extracting attributes that are invariant under polarization transformations. The polarimetric signatures are represented in terms of the components of the Stokes vectors. Invariant algebra is then used to extract a set of signature-related attributes that are invariant under linear transformation of the Stokes vectors. Experimental results using polarimetric infrared signatures of a number of manmade and natural objects undergoing systematic linear transformations support the invariancy of these attributes.

  5. Transformation invariant sparse coding

    DEFF Research Database (Denmark)

    Mørup, Morten; Schmidt, Mikkel Nørgaard

    2011-01-01

    Sparse coding is a well established principle for unsupervised learning. Traditionally, features are extracted in sparse coding in specific locations, however, often we would prefer invariant representation. This paper introduces a general transformation invariant sparse coding (TISC) model....... The model decomposes images into features invariant to location and general transformation by a set of specified operators as well as a sparse coding matrix indicating where and to what degree in the original image these features are present. The TISC model is in general overcomplete and we therefore invoke...... sparse coding to estimate its parameters. We demonstrate how the model can correctly identify components of non-trivial artificial as well as real image data. Thus, the model is capable of reducing feature redundancies in terms of pre-specified transformations improving the component identification....

  6. Plastid transformation in eggplant.

    Science.gov (United States)

    Bansal, Kailash C; Singh, Ajay K

    2014-01-01

    Eggplant (Solanum melongena L.) is an important vegetable crop of tropical and temperate regions of the world. Here we describe a procedure for eggplant plastid transformation, which involves preparation of explants, biolistic delivery of plastid transformation vector into green stem segments, selection procedure, and identification of the transplastomic plants. Shoot buds appear from cut ends of the stem explants following 5-6 weeks of spectinomycin selection after bombardment with the plastid transformation vector containing aadA gene as selectable marker. Transplastomic lines are obtained after the regenerated shoots are subjected to several rounds of spectinomycin selection over a period of 9 weeks. Homoplasmic transplastomic lines are further confirmed by spectinomycin and streptomycin double selection. The transplastomic technology development in this plant species will open up exciting possibilities for improving crop performance, metabolic engineering, and the use of plants as factories for producing biopharmaceuticals.

  7. Integrated magnetic transformer assembly

    DEFF Research Database (Denmark)

    2014-01-01

    The present invention relates to an integrated magnetics transformer assembly comprising a first magnetically permeable core forming a first substantially closed magnetic flux path and a second magnetically permeable core forming a second substantially closed magnetic flux path. A first input...... inductor winding is wound around a first predetermined segment of the first magnetically permeable core and a second input inductor winding is wound around a first predetermined segment of the second magnetically permeable core. The integrated magnetics transformer assembly further comprises a first output......-winding of the first output inductor winding and the first half-winding of the second output inductor winding are configured to produce aligned, i.e. in the same direction, magnetic fluxes through the first substantially closed magnetic flux path. The integrated magnetics transformer assembly is well- suited for use...

  8. Melanoma genetics

    DEFF Research Database (Denmark)

    Read, Jazlyn; Wadt, Karin A W; Hayward, Nicholas K

    2016-01-01

    Approximately 10% of melanoma cases report a relative affected with melanoma, and a positive family history is associated with an increased risk of developing melanoma. Although the majority of genetic alterations associated with melanoma development are somatic, the underlying presence...... of heritable melanoma risk genes is an important component of disease occurrence. Susceptibility for some families is due to mutation in one of the known high penetrance melanoma predisposition genes: CDKN2A, CDK4, BAP1, POT1, ACD, TERF2IP and TERT. However, despite such mutations being implicated...... in a combined total of approximately 50% of familial melanoma cases, the underlying genetic basis is unexplained for the remainder of high-density melanoma families. Aside from the possibility of extremely rare mutations in a few additional high penetrance genes yet to be discovered, this suggests a likely...

  9. Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment.

    Science.gov (United States)

    Ueki, Shoko; Lacroix, Benoît; Krichevsky, Alexander; Lazarowitz, Sondra G; Citovsky, Vitaly

    2009-01-01

    Transient gene expression is an indispensable tool for studying functions of gene products. In the case of plants, transient introduction of genes by Agrobacterium infiltration is a method of choice for many species. However, this technique does not work efficiently in Arabidopsis leaf tissue, the most widely used model system for basic plant biology research. Here we present an optimized protocol for biolistic delivery of plasmid DNA into the epidermis of Arabidopsis leaves, which can be easily performed using the Bio-Rad Helios gene gun system. This protocol yields efficient and reproducible transient expression of diverse genes and is exemplified here for use in a functional assay of a transcription repressor and for the subcellular localization and cell-to-cell movement of plant viral movement protein. This protocol is suitable for studies of biological function and subcellular localization of the gene product of interest directly in planta by utilizing different types of activity-based assays. Using this procedure, the data are obtained after 2-4 d of work.

  10. Genetic transformation of cry1EC gene into cotton (Gossypium ...

    African Journals Online (AJOL)

    welcome

    2013-04-10

    Apr 10, 2013 ... sp is not susceptible to Bt cotton containing Cry1Ac toxin so, there is ... encodes an insecticidal protein Cry1EC, (ii) 35S promoter from cauliflower ... the embryogenic calli were treated with the bacterial suspension by shaking ...

  11. Genetic transformation of lettuce (Lactuca sativa): A review

    African Journals Online (AJOL)

    SAM

    2014-04-16

    Apr 16, 2014 ... on using lettuce bioreactor to produce pharmaceutical protein and vaccines, ... Lettuce used as a foreign protein expression system has the .... Cholera toxin ..... sativa L.) with vectors bearing genes of bacterial antigenes from.

  12. Prochlorococcus Genetic Transformation and the Genomics of Nitrogen Metabolism

    Science.gov (United States)

    2005-09-01

    J. Structure and mutation ofa gene - USA (S.L.S.) encoding a M, 33000 phycocyanin -associated linker polypeptide. Arch. Microbiol. 153, 541-549 (1990...numerically 23. Dolganov, N. & Grossman, A. R. A polypeptide with similarity to phycocyanin alpha-subunit dominates the phytoplankton in the tropical and

  13. Studies on Agrobacterium-mediated genetic transformation of ...

    African Journals Online (AJOL)

    SERVER

    2008-03-04

    Mar 4, 2008 ... phosphotransferase (NPT II) gene. The presence of the SBD2 gene in the genomic DNA of transgenic ... accessible by direct sexual hybridization. A highly promising ..... bioassay with tobacco tissue cultures. Physiol. Plant.

  14. Similarity transformations of MAPs

    Directory of Open Access Journals (Sweden)

    Andersen Allan T.

    1999-01-01

    Full Text Available We introduce the notion of similar Markovian Arrival Processes (MAPs and show that the event stationary point processes related to two similar MAPs are stochastically equivalent. This holds true for the time stationary point processes too. We show that several well known stochastical equivalences as e.g. that between the H 2 renewal process and the Interrupted Poisson Process (IPP can be expressed by the similarity transformations of MAPs. In the appendix the valid region of similarity transformations for two-state MAPs is characterized.

  15. Transformation of technical infrastructure

    DEFF Research Database (Denmark)

    Nielsen, Susanne Balslev

    The scope of the project is to investigate the possibillities of - and the barriers for a transformation of technical infrastructure conserning energy, water and waste. It focus on urban ecology as a transformation strategy. The theoretical background of the project is theories about infrastructure......, the evolution of large technological systems and theories about organisational and technological transformationprocesses. The empirical work consist of three analysis at three different levels: socio-technical descriptions of each sector, an envestigation of one municipality and envestigations of one workshop...

  16. Transformer room fire tests

    Science.gov (United States)

    Fustich, C. D.

    1980-03-01

    A series of transformer room fire tests are reported to demonstate the shock hazard present when automatic sprinklers operate over energized electrical equipment. Fire protection was provided by standard 0.5 inch pendent automatic sprinklers temperature rated at 135 F and installed to give approximately 150 sq ft per head coverage. A 480 v dry transformer was used in the room to provide a three phase, four wire distribution system. It is shown that the induced currents in the test room during the various tests are relatively small and pose no appreciable personnel shock hazard.

  17. Studying Voltage Transformer Ferroresonance

    Directory of Open Access Journals (Sweden)

    Hamid Radmanesh

    2012-09-01

    Full Text Available This study studies the effect of Circuit Breaker Shunt Resistance (CBSR, Metal Oxide Vaistor (MOV and Neutral earth Resistance (NR on the control of ferroresonance in the voltage transformer. It is expected that NR can controlled ferroresonance better than MOV and CBSR. Study has been done on a one phase voltage transformer rated 100 VA, 275 kV. The simulation results reveal that considering the CBSR and MOV exhibits a great mitigating effect on ferroresonance overvoltages, but these resistances cannot control these phenomena for all range of parameters. By applying NR to the system structure, ferroresonance has been controlled and its amplitude has been damped for all parameters values.

  18. Displacive Transformation in Ceramics

    Science.gov (United States)

    1994-02-28

    single cycle[21]. In zirconia , ferroelastic domains appeared during the cubic to tetragonal transformation at -2200’C, where [c] axes were elongated...Mechanism in Tetragonal Zirconia ( TZP ) Ceramics," Adv. in Ceramics 24 (1986) 653-662. 26. K. Mehta, J. F. Jue and A. V. Virkar, "Grinding-Liduced...barium copper oxide (YBa2Cu306+x) and dicalcium silicate (Ca 2 SiO4 ). The cubic to tetragonal transformation in PbTiO3 40 was proven to be

  19. The quantum Arnold transformation

    Science.gov (United States)

    Aldaya, V.; Cossío, F.; Guerrero, J.; López-Ruiz, F. F.

    2011-02-01

    Using a quantum version of the Arnold transformation of classical mechanics, all quantum dynamical systems whose classical equations of motion are non-homogeneous linear second-order ordinary differential equations, including systems with friction linear in velocity, can be related to the quantum free-particle dynamical system. This transformation provides a basic (Heisenberg-Weyl) algebra of quantum operators, along with well-defined Hermitian operators which can be chosen as evolution-like observables and complete the entire Schrödinger algebra. It also proves to be very helpful in performing certain computations quickly, to obtain, for example, wavefunctions and closed analytic expressions for time-evolution operators.

  20. Specificity of Czech Economic Transformation

    OpenAIRE

    Haberle, Ondřej

    2010-01-01

    The aim of this thesis is to describe the economic transformation in Czech Republic in 1990-1997 with accent on difference between transformation process in Czech Republic and other transition countries. Theoretical part is based on the description of the model of centrally planned economy and transformation theory. Practical part discuss macroeconomy a microeconomy measures of economic transformation.

  1. Genetic Engineering of Alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Wang, Dan; Khurshid, Muhammad; Sun, Zhan Min; Tang, Yi Xiong; Zhou, Mei Liang; Wu, Yan Min

    2016-01-01

    Alfalfa is excellent perennial legume forage for its extensive ecological adaptability, high nutrition value, palatability and biological nitrogen fixation. It plays a very important role in the agriculture, animal husbandry and ecological construction. It is cultivated in all continents. With the development of modern plant breeding and genetic engineering techniques, a large amount of work has been carried out on alfalfa. Here we summarize the recent research advances in genetic engineering of alfalfa breeding, including transformation, quality improvement, stress resistance and as a bioreactor. The review article can enables us to understand the research method, direction and achievements of genetic engineering technology of Alfalfa.

  2. Melanocyte Transformation Associated with Substrate Adhesion Impediment

    Directory of Open Access Journals (Sweden)

    Sueli M. Oba-Shinjo

    2006-03-01

    Full Text Available Exclude experimental models of malignant transformation employ chemical and physical carcinogens or genetic manipulations to study tumor progression. In this work, different melanoma cell lines were established after submitting a nontumorigenic melanocyte lineage (melan-a to sequential cycles of forced anchorage impediment. The great majority of these cells underwent anoikis when maintained in suspension. After one deadhesion cycle, phenotypic alterations were noticeable in the few surviving cells, which became more numerous and showed progressive alterations after each adhesion impediment step. No significant differences in cell surface expression of integrins were detected, but a clear electrophoretic migration shift, compatible with an altered glycosylation pattern, was observed for β1 chain in transformed cell lines. In parallel, a progressive enrichment of tri- and tetra-antennary N-glycans was apparent, suggesting increased N-acetylglucosaminyl-transferase V activity. Alterations both in proteoglycan glycosylation pattern and core protein expression were detected during the transformation process. In conclusion, this model corroborates the role of adhesion state as a promoting agent in transformation process and demonstrates that cell adhesion disturbances may act as carcinogenic stimuli, at least for a nontumorigenic immortalized melanocyte lineage. These findings have intriguing implications for in vivo carcinogenesis, suggesting that anchorage independence may precede, and contribute to, neoplastic conversion.

  3. 利用草甘膦作为大豆子叶节遗传转化筛选剂的可行性分析%Feasibility Analysis of Using Glyphosate as the Selective Agent in Soybean Genetic Transformation System with Cotyledonary Nodes as Explants

    Institute of Scientific and Technical Information of China (English)

    岳岩磊; 于丽杰; 孙石; 韩天富; 侯文胜

    2012-01-01

    以4个非转基因大豆品种自贡冬豆、吉林小粒1号、中黄30和Williams 82以及2个抗草甘膦转基因大豆品系中作J9331和中作J9333为材料,利用大豆子叶节进行遗传转化,草甘膦作为筛选剂,以丛生芽诱导率、成活率和伸长率为考察指标,探索丛生芽诱导期、筛选期和伸长期的适宜草甘膦筛选浓度.结果表明,丛生芽诱导期、筛选期及伸长期的草甘膦适宜筛选浓度范围分别为100~160、220 ~ 240和5~ 10 mg·L-1,3个时期最适宜筛选浓度分别为60、240和5 mg·L-1.%Using four conventional soybean varieties ( Zigongdongdou, Jilinxiaoli-1 , Zhonghuang 30 and Williams 82) and two glyphosate-resistant transgenic soybean lines ( Zhongzuo J9331 and J9333) ,the optimal concentrations of glyphosate were explored at three stages of soybean genetic transformation system with the cotyledon nodes as explants. The results showed that it is feasible to use glyphosate as the selective agent,the suitable concentrations are 100-160,220-240 and 5-10 mg·L-1 respectively at the stages of the induction, screening and elongation of multiple shoots clumps. In the whole, the optimal screening effect can be gotten with the glyphosate concentrotions as 60,240 and 5 mg · L-1 respectively at the three stages.

  4. Improved injection needles facilitate germline transformation of the buckeye butterfly Junonia coenia.

    Science.gov (United States)

    Beaudette, Kahlia; Hughes, Tia M; Marcus, Jeffrey M

    2014-01-01

    Germline transformation with transposon vectors is an important tool for insect genetics, but progress in developing transformation protocols for butterflies has been limited by high post-injection ova mortality. Here we present an improved glass injection needle design for injecting butterfly ova that increases survival in three Nymphalid butterfly species. Using the needles to genetically transform the common buckeye butterfly Junonia coenia, the hatch rate for injected Junonia ova was 21.7%, the transformation rate was 3%, and the overall experimental efficiency was 0.327%, a substantial improvement over previous results in other butterfly species. Improved needle design and a higher efficiency of transformation should permit the deployment of transposon-based genetic tools in a broad range of less fecund lepidopteran species.

  5. Genetic Testing for ALS

    Science.gov (United States)

    ... Involved Donate Familial Amyotrophic Lateral Sclerosis (FALS) and Genetic Testing By Deborah Hartzfeld, MS, CGC, Certified Genetic Counselor ... in your area, please visit www.nsgc.org . Genetic Testing Genetic testing can help determine the cause of ...

  6. Genetic Science Learning Center

    Science.gov (United States)

    ... Mouse Party on Learn.Genetics.utah.edu Students doing the Tree of Genetic Traits activity Learn.Genetics is one of the most widely used science education websites in the world The Community Genetics ...

  7. Efficient transformation of Mycosphaerella fijiensis by underwater shock waves.

    Science.gov (United States)

    Escobar-Tovar, Lina; Magaña-Ortíz, Denis; Fernández, Francisco; Guzmán-Quesada, Mauricio; Sandoval-Fernández, Jorge A; Ortíz-Vázquez, Elizabeth; Loske, Achim M; Gómez-Lim, Miguel A

    2015-12-01

    Black leaf streak disease, also known as black Sigatoka, causes dramatic losses in production of banana and plantains fruits. The disease is caused by the pathogenic fungus Mycosphaerella fijiensis (anamorph Pseudocercospora fijiensis; Mycosphaerellaceae). Genetic transformation of M. fijiensis would allow a better understanding of molecular basis of pathogenicity and design novel approaches to control the infection caused by this pathogen. However, transformation of this fungus has not been easy. We report here a protocol for genetic transformation of M. fijiensis employing underwater shock waves and intact conidia. The recombinant strains recovered showed genetic stability over >10 generations. The frequency of transformation obtained was between 75 and 150 times higher than the efficiency reported in the only article published on transformation of M. fijiensis using spheroplasts. This improvement allowed the use of a thousand times less cells than the amount employed before, avoiding the need for cumbersome successive batch cultures. Our protocol is simple, highly efficient, fast and reproducible and together with the available genomes of M. fijiensis and Musa acuminata, it offers new possibilities to study the diverse mechanisms of pathogenesis of the fungus. Copyright © 2015. Published by Elsevier B.V.

  8. Fourier transforms principles and applications

    CERN Document Server

    Hansen, Eric W

    2014-01-01

    Fourier Transforms: Principles and Applications explains transform methods and their applications to electrical systems from circuits, antennas, and signal processors-ably guiding readers from vector space concepts through the Discrete Fourier Transform (DFT), Fourier series, and Fourier transform to other related transform methods.  Featuring chapter end summaries of key results, over two hundred examples and four hundred homework problems, and a Solutions Manual this book is perfect for graduate students in signal processing and communications as well as practicing engineers.

  9. Genetic circuit design automation.

    Science.gov (United States)

    Nielsen, Alec A K; Der, Bryan S; Shin, Jonghyeon; Vaidyanathan, Prashant; Paralanov, Vanya; Strychalski, Elizabeth A; Ross, David; Densmore, Douglas; Voigt, Christopher A

    2016-04-01

    Computation can be performed in living cells by DNA-encoded circuits that process sensory information and control biological functions. Their construction is time-intensive, requiring manual part assembly and balancing of regulator expression. We describe a design environment, Cello, in which a user writes Verilog code that is automatically transformed into a DNA sequence. Algorithms build a circuit diagram, assign and connect gates, and simulate performance. Reliable circuit design requires the insulation of gates from genetic context, so that they function identically when used in different circuits. We used Cello to design 60 circuits forEscherichia coli(880,000 base pairs of DNA), for which each DNA sequence was built as predicted by the software with no additional tuning. Of these, 45 circuits performed correctly in every output state (up to 10 regulators and 55 parts), and across all circuits 92% of the output states functioned as predicted. Design automation simplifies the incorporation of genetic circuits into biotechnology projects that require decision-making, control, sensing, or spatial organization.

  10. Transformative Mixed Methods Research

    Science.gov (United States)

    Mertens, Donna M.

    2010-01-01

    Paradigms serve as metaphysical frameworks that guide researchers in the identification and clarification of their beliefs with regard to ethics, reality, knowledge, and methodology. The transformative paradigm is explained and illustrated as a framework for researchers who place a priority on social justice and the furtherance of human rights.…

  11. On the Meijer transformation

    Directory of Open Access Journals (Sweden)

    J. Conlan

    1978-01-01

    Full Text Available Recently [8], an operational calculus for the operator Bμ=t−μDt1+μD with −1<μ<∞ was developed via the algebraic approach [4], [13], [15]. This paper gives the integral transform version. In particular, a differentiation theorem and a convolution theorem are proved.

  12. Transforming Military Leaders

    Science.gov (United States)

    2006-03-15

    McGraw Hill, 2005), 608. 41 Peter Guy Northouse , Leadership : Theory and Practice (New York: Sage Publications Inc, 2004), 151. 42 Ibid, 151. 43...KEY TERMS: Globalization, Military Transformation, Leadership CLASSIFICATION: Unclassified A phrase that captures the challenges associated with the... leadership competencies to fully execute our National Security Strategy across the parochial elements of diplomatic, information, military and economic

  13. Transforming Public Space

    DEFF Research Database (Denmark)

    Navarro, Dora

    2009-01-01

    , such as municipal elections and the district's participatory budget. In addition, I show how the public uses the radio to channel their claims. I also identify the factors that prevent the radio from fully empowering the public and transforming public space into a more critical and democratic one....

  14. Transformative Mixed Methods Research

    Science.gov (United States)

    Mertens, Donna M.

    2010-01-01

    Paradigms serve as metaphysical frameworks that guide researchers in the identification and clarification of their beliefs with regard to ethics, reality, knowledge, and methodology. The transformative paradigm is explained and illustrated as a framework for researchers who place a priority on social justice and the furtherance of human rights.…

  15. Military Culture and Transformation

    Science.gov (United States)

    2008-01-01

    mines the success of transformation. The period between 1914 and 1945 shows the dynamic nature of military innova - tion and the difficulty military...maximized unity of effort. Leaders can foster a disciplined culture that encourages change and innova - tion by “creating a consistent system with clear

  16. Graph Transforming Java Data

    NARCIS (Netherlands)

    Mol, de Maarten; Rensink, Arend; Hunt, James J.

    2012-01-01

    This paper introduces an approach for adding graph transformation-based functionality to existing JAVA programs. The approach relies on a set of annotations to identify the intended graph structure, as well as on user methods to manipulate that structure, within the user’s own JAVA class declaration

  17. Transformations, Dynamics and Complexity

    CERN Document Server

    Glazunov, Nikolaj

    2011-01-01

    We review and investigate some new problems and results in the field of dynamical systems generated by iteration of maps, {\\beta}-transformations, partitions, group actions, bundle dynamical systems, Hasse-Kloosterman maps, and some aspects of complexity of the systems.

  18. Lost in transformation?

    DEFF Research Database (Denmark)

    Norlyk, Annelise; Haahr, Anita; Dreyer, Pia

    2017-01-01

    and values from evidence-based medicine are being lost in the transformation into the current evidence-based hospital culture which potentially leads to a McDonaldization of nursing practice reflected as ‘one best way’. We argue for reviving ethics of care perspectives in today’s evidence practice...

  19. Transformer Impedance Reflection Demonstration

    Science.gov (United States)

    Layton, William

    2014-01-01

    Questions often arise as to how a device attached to a transformer can draw power from the electrical power grid since it seems that the primary and secondary are not connected to one another. However, a closer look at how the primary and secondary are linked together magnetically and a consideration of the role of Lenz's law in this linkage…

  20. Transformer and Meter Tester

    Science.gov (United States)

    Stoms, R. M.

    1984-01-01

    Numerically-controlled 5-axis machine tool uses transformer and meter to determine and indicate whether tool is in home position, but lacks built-in test mode to check them. Tester makes possible test, and repair of components at machine rather then replace them when operation seems suspect.