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Vanamala Anjaiah - One of the best experts on this subject based on the ideXlab platform.
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an efficient method for the production of Transgenic Plants of peanut arachis hypogaea l through agrobacterium tumefaciens mediated genetic transformation
Plant Science, 2000Co-Authors: Kiran K Sharma, Vanamala AnjaiahAbstract:Cotyledon exPlants from mature peanut seeds (Arachis hypogaea L.) were optimized to obtain adventitious shoot buds with high frequencies (>90%). Efficient transformation of these cotyledons by using Agrobacterium tumefaciens strain C58 carrying neomycin phosphotransferase II (nptII) and s-glucuronidase (GUS; uidA), or coat protein gene of the Indian peanut clump virus (IPCVcp) and nptII on binary vectors (pBI121; pROKII:IPCVcp) led to the production of a large percentage (55%) of Transgenic Plants. Transformed individuals were obtained through selection on medium containing 125 mg l(-1) kanamycin. A large number of independently transformed Plants (over 75) were successfully transplanted to the glasshouse. Integration of the transgenes and stable genetic transformants in the progeny were assessed by PCR amplification of 700-bp fragment of nptII and 585-bp of IPCVcp genes, and Southern blot hybridizations in the T1 generation of Transgenic Plants. Analysis of 35 Transgenic Plants of T1 generation from the progeny of a single transformation event suggested the segregation of a single copy insert in a 3:1 Mendelian ratio. On an average, 120-150 days were required between the initiation of explant transformation and transfer of rooted Plants to the greenhouse. The cotyledon regeneration system proved to be an excellent vehicle for the production of a large number of independently transformed peanut Plants. Shoot formation was rapid and prolific, and a large proportion of these shoots developed into fertile Plants. The method reported here provides new opportunities for the crop improvement of peanut via genetic transformation.
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an efficient method for the production of Transgenic Plants of peanut arachis hypogaea l through agrobacterium tumefaciens mediated genetic transformation
Plant Science, 2000Co-Authors: Kiran K Sharma, Vanamala AnjaiahAbstract:Cotyledon exPlants from mature peanut seeds (Arachis hypogaea L.) were optimized to obtain adventitious shoot buds with high frequencies (\90%). Efficient transformation of these cotyledons by using Agrobacterium tumefaciens strain C58 carrying neomycin phosphotransferase II (nptII) and s-glucuronidase (GUS; uidA), or coat protein gene of the Indian peanut clump virus (IPCVcp) and nptII on binary vectors (pBI121; pROKII:IPCVcp) led to the production of a large percentage (55%) of Transgenic Plants. Transformed individuals were obtained through selection on medium containing 125 mg l�1 kanamycin. A large number of independently transformed Plants (over 75) were successfully transplanted to the glasshouse. Integration of the transgenes and stable genetic transformants in the progeny were assessed by PCR amplification of 700-bp fragment of nptII and 585-bp of IPCVcp genes, and Southern blot hybridizations in the T1 generation of Transgenic Plants. Analysis of 35 Transgenic Plants of T1 generation from the progeny of a single transformation event suggested the segregation of a single copy insert in a 3:1 Mendelian ratio. On an average, 120–150 days were required between the initiation of explant transformation and transfer of rooted Plants to the greenhouse. The cotyledon regeneration system proved to be an excellent vehicle for the production of a large number of independently transformed peanut Plants. Shoot formation was rapid and prolific, and a large proportion of these shoots developed into fertile Plants. The method reported here provides new opportunities for the crop improvement of peanut via genetic transformation. © 2000 Elsevier Science Ireland Ltd. All rights reserved.
Kiran K Sharma - One of the best experts on this subject based on the ideXlab platform.
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an efficient method for the production of Transgenic Plants of peanut arachis hypogaea l through agrobacterium tumefaciens mediated genetic transformation
Plant Science, 2000Co-Authors: Kiran K Sharma, Vanamala AnjaiahAbstract:Cotyledon exPlants from mature peanut seeds (Arachis hypogaea L.) were optimized to obtain adventitious shoot buds with high frequencies (>90%). Efficient transformation of these cotyledons by using Agrobacterium tumefaciens strain C58 carrying neomycin phosphotransferase II (nptII) and s-glucuronidase (GUS; uidA), or coat protein gene of the Indian peanut clump virus (IPCVcp) and nptII on binary vectors (pBI121; pROKII:IPCVcp) led to the production of a large percentage (55%) of Transgenic Plants. Transformed individuals were obtained through selection on medium containing 125 mg l(-1) kanamycin. A large number of independently transformed Plants (over 75) were successfully transplanted to the glasshouse. Integration of the transgenes and stable genetic transformants in the progeny were assessed by PCR amplification of 700-bp fragment of nptII and 585-bp of IPCVcp genes, and Southern blot hybridizations in the T1 generation of Transgenic Plants. Analysis of 35 Transgenic Plants of T1 generation from the progeny of a single transformation event suggested the segregation of a single copy insert in a 3:1 Mendelian ratio. On an average, 120-150 days were required between the initiation of explant transformation and transfer of rooted Plants to the greenhouse. The cotyledon regeneration system proved to be an excellent vehicle for the production of a large number of independently transformed peanut Plants. Shoot formation was rapid and prolific, and a large proportion of these shoots developed into fertile Plants. The method reported here provides new opportunities for the crop improvement of peanut via genetic transformation.
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an efficient method for the production of Transgenic Plants of peanut arachis hypogaea l through agrobacterium tumefaciens mediated genetic transformation
Plant Science, 2000Co-Authors: Kiran K Sharma, Vanamala AnjaiahAbstract:Cotyledon exPlants from mature peanut seeds (Arachis hypogaea L.) were optimized to obtain adventitious shoot buds with high frequencies (\90%). Efficient transformation of these cotyledons by using Agrobacterium tumefaciens strain C58 carrying neomycin phosphotransferase II (nptII) and s-glucuronidase (GUS; uidA), or coat protein gene of the Indian peanut clump virus (IPCVcp) and nptII on binary vectors (pBI121; pROKII:IPCVcp) led to the production of a large percentage (55%) of Transgenic Plants. Transformed individuals were obtained through selection on medium containing 125 mg l�1 kanamycin. A large number of independently transformed Plants (over 75) were successfully transplanted to the glasshouse. Integration of the transgenes and stable genetic transformants in the progeny were assessed by PCR amplification of 700-bp fragment of nptII and 585-bp of IPCVcp genes, and Southern blot hybridizations in the T1 generation of Transgenic Plants. Analysis of 35 Transgenic Plants of T1 generation from the progeny of a single transformation event suggested the segregation of a single copy insert in a 3:1 Mendelian ratio. On an average, 120–150 days were required between the initiation of explant transformation and transfer of rooted Plants to the greenhouse. The cotyledon regeneration system proved to be an excellent vehicle for the production of a large number of independently transformed peanut Plants. Shoot formation was rapid and prolific, and a large proportion of these shoots developed into fertile Plants. The method reported here provides new opportunities for the crop improvement of peanut via genetic transformation. © 2000 Elsevier Science Ireland Ltd. All rights reserved.
Ayalew Mentewab - One of the best experts on this subject based on the ideXlab platform.
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overexpression of an arabidopsis thaliana abc transporter confers kanamycin resistance to Transgenic Plants
Nature Biotechnology, 2005Co-Authors: Ayalew Mentewab, Neal C StewartAbstract:Overexpression of an Arabidopsis thaliana ABC transporter confers kanamycin resistance to Transgenic Plants
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Overexpression of an Arabidopsis thaliana ABC transporter confers kanamycin resistance to Transgenic Plants
Nature biotechnology, 2005Co-Authors: Ayalew Mentewab, C. Neal StewartAbstract:Selectable markers of bacterial origin such as the neomycin phosphotransferase type II gene, which can confer kanamycin resistance to Transgenic Plants, represent an invaluable tool for plant engineering. However, since all currently used antibiotic-resistance genes are of bacterial origin, there have been concerns about horizontal gene transfer from Transgenic Plants back to bacteria, which may result in antibiotic resistance. Here we characterize a plant gene, Atwbc19, the gene that encodes an Arabidopsis thaliana ATP binding cassette (ABC) transporter and confers antibiotic resistance to Transgenic Plants. The mechanism of resistance is novel, and the levels of resistance achieved are comparable to those attained through expression of bacterial antibiotic-resistance genes in Transgenic tobacco using the CaMV 35S promoter. Because ABC transporters are endogenous to Plants, the use of Atwbc19 as a selectable marker in Transgenic Plants may provide a practical alternative to current bacterial marker genes in terms of the risk for horizontal transfer of resistance genes.
Andres Wigdorovitz - One of the best experts on this subject based on the ideXlab platform.
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protective lactogenic immunity conferred by an edible peptide vaccine to bovine rotavirus produced in Transgenic Plants
Journal of General Virology, 2004Co-Authors: Andres Wigdorovitz, Marina Valeria Mozgovoj, Maria Jose Dus Santos, Viviana Parreno, Cristina Gomez, Daniel M Perezfilgueira, K Trono, Raul Rios, Pascual M FranzoneAbstract:Vaccines produced in Transgenic Plants constitute a promising alternative to conventional immunogens, presenting the possibility of stimulating secretory and systemic immunity against enteric pathogens when administered orally. Protection against enteric pathogens affecting newborn animals requires, in most cases, the stimulation of lactogenic immunity. Here, the group presents the development of an experimental immunogen based on expression of an immunorelevant peptide, eBRV4, of the VP4 protein of bovine rotavirus (BRV), which has been described as harbouring at least one neutralizing epitope as well as being responsible for the adsorption of the virus to epithelial cells. The eBRV4 epitope was efficiently expressed in Transgenic alfalfa as a translational fusion protein with the highly stable reporter enzyme β-glucuronidase (βGUS), which served as a carrier, stabilized the synthesized peptide and facilitated screening for the higher expression levels in Plants. Correlation of expression of the eBRV4 epitope in Plants with those presenting the highest βGUS activities was confirmed by a Western blot assay specific for the BRV peptide. The eBRV4 epitope expressed in Plants was effective in inducing an anti-rotavirus antibody response in adult female mice when administered either intraperitoneally or orally and, more importantly, suckling mice born from immunized female mice were protected against oral challenge with virulent rotavirus. These results demonstrate the feasibility of inducing lactogenic immunity against an enteric pathogen using an edible vaccine produced in Transgenic Plants.
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high yield expression of a viral peptide vaccine in Transgenic Plants
FEBS Letters, 2001Co-Authors: Alejandro Brun, Andres Wigdorovitz, Rafael Catala, Jorge L Martineztorrecuadrada, Ignacio Casal, Julio Salinas, M V Borca, Jose M EscribanoAbstract:A high-yield production of a peptide vaccine in Transgenic Plants is described here. A 21-mer peptide, which confers protection to dogs against challenge with virulent canine parvovirus, has been expressed in Transgenic Plants as an amino-terminal translational fusion with the GUS gene. Transformants were selected on the basis of their GUS activities, showing expression levels of the recombinant protein up to 3% of the total leaf soluble protein, a production yield comparable to that obtained with the same epitope expressed by chimeric plant viruses. The immunogenicity of the plant-derived peptide was demonstrated in mice immunized either intraperitoneally or orally with Transgenic plant extracts, providing the suitability of the GUS fusions approach for low-cost production of peptide vaccines.
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induction of a protective antibody response to foot and mouth disease virus in mice following oral or parenteral immunization with alfalfa Transgenic Plants expressing the viral structural protein vp1
Virology, 1999Co-Authors: Andres Wigdorovitz, K Trono, Raul Rios, Pascual M Franzone, C Carrillo, M Dus J Santos, Andrea Peralta, Maria Cristina Gomez, A M Sadir, Jose Angel Martinez EscribanoAbstract:The utilization of Transgenic Plants expressing recombinant antigens to be used in the formulation of experimental immunogens has been recently communicated. We report here the development of Transgenic Plants of alfalfa expressing the structural protein VP1 of foot and mouth disease virus (FMDV). The presence of the transgenes in the Plants was confirmed by PCR and their specific transcription was demonstrated by RT-PCR. Mice parenterally immunized using leaf extracts or receiving in their diet freshly harvested leaves from the Transgenic Plants developed a virus-specific immune response. Animals immunized by either method elicited a specific antibody response to a synthetic peptide representing amino acid residues 135-160 of VP1, to the structural protein VP1, and to intact FMDV particles. Additionally, the immunized mice were protected against experimental challenge with the virus. We believe this is the first report demonstrating the induction of a protective systemic antibody response in animals fed Transgenic Plants expressing a viral antigen. These results support the feasibility of producing edible vaccines in Transgenic forage Plants, such as alfalfa, commonly used in the diet of domestic animals even for those antigens for which a systemic immune response is required.
Neal C Stewart - One of the best experts on this subject based on the ideXlab platform.
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overexpression of an arabidopsis thaliana abc transporter confers kanamycin resistance to Transgenic Plants
Nature Biotechnology, 2005Co-Authors: Ayalew Mentewab, Neal C StewartAbstract:Overexpression of an Arabidopsis thaliana ABC transporter confers kanamycin resistance to Transgenic Plants
