Stable genetic transformation in cotton (Gossypium hirsutum L.) using marker genes

Article history: Received 24 October 2013 Revised form 29 October 2013 Accepted 3 November 2013 Available online 25 January 2014 Transformation of novel candidate gene for cotton fiber improvement is using a unique Agrobacterium Mediated Transformation approach. The detection of NPTII (kanamycin resistance) as a transgenic marker gene and 35SCaMV as promoter in cotton has been carried out. Transgenic cotton plants were also confirmed by using VirG gene of Agrobacterium. Genomic DNA from transgenic cotton samples were isolated using Fermentas DNA extraction kit. The purity of DNA samples was estimated using UV-Vis spectrophotometry at 260 nm and 280 nm. They gave an absorbance ratio (A260/A280) of 1.74-1.79 which indicated its purities. In order to identify the transgenic cotton, a 151bp fragment of the NPTII gene (kanamycin resistant gene), 210bp fragment of 35SCaMV promoter and 410bp of VirG gene of Agrobacterium were amplified using polymerase chain reaction (PCR). The results showed that both NPTII gene and 35SCaMV promoter are present in putative transgenic cotton plants but no amplification was found in untransformed plant. 2013 KnowledgesPublisher Ltd. All rights reserved.