Cloning of Bowman-Birk Type Protease Inhibitor Gene from Pulse Crops Datta S1,*, Tiwari Kapil K2 1Biotechnology Unit, Division of Crop Improvement, Indian Institute of Pulses Research, Kanpur-208024, India 2Department of Agricultural Biotechnology, C S Azad University of Agriculture & Technology, Kanpur-208002, India *Corresponding author: (E-mail: subhojit@email.com)
Abstract Plant derived insecticidal genes are increasingly being used to develop insect resistant transgenic crops. In order to clone of protease inhibitor genes from legumes, 18 to 23 bp long sucleotides primers were designed using sequence database of public domain. PCR amplifications of selected genotypes were carried out at different annealing temperatures. In case of BBPI-II primer, 42°c annealing temperature resulted in amplification of 650 bp single fragment in lentil varieties IPL 81 and DPL15. Isolated 650bp fragment was ligated to linearised vector pTZ57R/T. After ligation, recombinant vector was transformed in bacterial cell of E. coli strain JM 109. Top Key words Protease inhibitor, cloning, pulse crops. Top |