Cloning of cry1Ac Gene from A New Indigenous Isolate of Bacillus thuringiensis

The present study was carried out with the objective of characterization and cloning of cry1Ac gene from indigenous isolate of Bacillus thuringiensis (Bt). Crystal inclusion study revealed that the isolate showed bipyramidal like and cuboidal inclusions. The protein profile data showed faint band of ∼135 kDa and strong ∼65 kDa protein bands. Further, screening of isolate of Bt by Helicoverpa armigera and Spodoptera litura bioassay showed different level of toxicity. PCR amplification was obtained with primers specific for cry1 family gene. Therefore, cry1 gene fragment(s) was amplified and cloned intop JET1.2/blunt cloning vector. Nucleotide data showed 99% homology to the sequence holotype of cry1Ac. Deduced amino acid sequence showed variation of single amino acid at 1076^th position of holotype Cry1Ac.