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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
In present investigation we performed Agrobacterium-mediated transformation in Coker-312 (Gossypium hirsutum L.). Freshly pre-cultured (48 hrs.) hypocotyl explants were infected with two different Agrobacterium tumifaciens strain EHA105 and LBA4404 carrying cry1Ac and cry1Acm respectively along with neomycin phosphotransferase - II gene as selection marker. Selection of kanamycin resistant embryogenic calli were performed on MS media supplemented 0.1 mg/l 2, 4-D + 0.5 mg/l kinetin with cefotaxime (400 mg/l) and kanamycin (100 mg/l), followed by subculture and maintenance for embryo maturation. Putative transformants were screened for the presence of specific gene and neomycin phosphotransferase (npt-II) by for its transgene amplification. Further, expression analysis was carried out using Reverse transcriptase-PCR (RT-PCR). Protein expression was checked with Bt expression strips in T0 generation. Further analysis of T1 plants demonstrated the failure detection of the transgene.