title: Development of insect-resistant transgenic cotton with chimeric TVip3A accumulating in chloroplasts.
abstract: An optimized vip3A gene, designated as vip3A* was chemically synthesized and a thi1 gene chloroplast transit peptide coding sequence was attached to its 5' end to produce the tvip3A*. vip3A* and tvip3A* genes were transformed into Gossypium hirsutum cv. Zhongmiansuo35 mediated by Agrobacterium tumefaciens. Four independent transgenic T1 lines with single-copy insertions and unchanged phenotypes (CTV1 and CTV2 for tvip3A*, and CV1 and CV2 for vip3A*) were selected by Polymerase chain reaction (PCR), Reverse transcription (RT)-PCR, Southern blotting, enzyme-linked immunosorbent assay (ELISA), and insect bioassay. As expected, the Vip3A* protein of CTV1 and CTV2 were transported to the chloroplasts, where they accumulated. Our results suggest that the two tvip3A* transgenic lines (CTV1 and CTV2) can be used to develop insect-resistant cultivars and could be used as a resource for raising multi-toxins-expressing transgenic cotton.
authors.affiliation: State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China. wujiahe@im.ac.cn
authors.forename: Jiahe
authors.initials: J
authors.lastname: Wu
authors.forename: Yingchuan
authors.initials: Y
authors.lastname: Tian
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D003368:Gossypium / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D010572:Pest Control, Biological / Q000379:methods*; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D036226:Seedlings / Q000254:growth & development; D018411:Spodoptera / Q000502:physiology*; D014158:Transcription, Genetic; D014170:Transformation, Genetic
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C447139:Vip3A protein, Bacillus thuringiensis