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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

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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 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2017.6(12): 2771-2779
DOI: https://doi.org/10.20546/ijcmas.2017.612.321


Agrobacterium tumefaciens Mediated Genetic Transformation in Coker-312 (Gossypium hirsutum L.) Using Hypocotyls Explants
M.P. Jadhav and I.S. Katageri*
Department of Biotechnology, University of Agricultural Sciences, Dharwad - 580 005, Karnataka, India
*Corresponding author
Abstract:

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.


Keywords: Agrobacterium, Genetic transformation, Gossypium hirsutum, RT-PCR, Strip ELISA.
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How to cite this article:

Jadhav, M.P. and Katageri, I.S. 2017. Agrobacterium tumefaciens Mediated Genetic Transformation in Coker-312 (Gossypium hirsutum L.) Using Hypocotyls Explants.Int.J.Curr.Microbiol.App.Sci. 6(12): 2771-2779. doi: https://doi.org/10.20546/ijcmas.2017.612.321