Abstract
The cryIII gene, isolated from a locally isolated Bacillus thuringiensis strain,CAMB #30382 (isolated from grain dust of Shakargarh,Punjab,Pakistan), highly effective against red flour beetle Tribolium castaneum, was amplified through Polymerase chain reaction (PCR) by using of specific primers, The cryIII gene was cloned in HindІІІ digested and dephosphorylated expression vector pHB201. For this purpose, the DNA was run through 0.8% agarose gel and was eluted by means of DNA extraction kit.The transformation was done into an acrystalliferous strain of Bacillus thuringiensis, 4D22, by electroporation method . The positive clones were selected onto the petri plates containing LB X-gal/IPTG/Chloramphenicol. The Bacillus thuringiensis containing the gene of interest were also confirmed through PCR, and restriction analysis. Total protein extract from the transformed and non transformed Bt. strains were collected through affinity column chromatography.
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