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)
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.
Protease inhibitor, cloning, pulse crops
