PCR Amplification and Cloning of Proteinase Inhibitor Gene in Cotton (Gossypium species)

The main objective of the work was to see presence of the three different Proteinase inhibitor genes of soybean Viz. Kti3, CI-II and PI-IV genes in cotton by synthesizing specific primer from already published data. DNA was extracted from different cotton spp. and was used for PCR amplification by the different specific primers for the Kti3, CI-II and PI-IV genes. Results have shown expected fragments of 651 bp for Kti3 gene and 250 bp for CI-II and PI-IV genes. These PCR fragments were were electroeluted from the Agarose gels and then used for cloning. These genes were then cloned in plasmid vector. Selection of transformed colony was done on the basis of Blue-White screnning. Plasmid was isolated and results were reconfirmed by doing PCR amplification of the Kti3, CI-II and PI-IV genes using specific primers. Results showed that the expected fragments were successfully cloned in the plasmid.