| TECHNIQUES AND METHODS |
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| Activity of a New Recombinant Insecticidal Crystal Protein Encoded by HJC Gene from Bacillus thuringiensis |
| SU Xu, WANG Jing, LV Li-kun, LI Li, YANG Dong-jing, LIU Hong-liang |
| Tianjin Center for Disease Control and Prevention, Tianjin 300011, China |
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Abstract For a better management of insect cross-resistance against Bt toxins, a fusion gene derived from cry1Ab and vip3 genes was created and named HJC. The gene was expressed in E. coli strain BL21 upon transformation with plasmid pET28a-HJC. The his-tagged HJC protein mainly existed as inclusion body in E. coli, and was purified by Ni-NTA resin affinity chromatography under denatured condition. After refolding upon gradient urea dialysis, recombinant HJC protein was analyzed for its immune recognition and insecticidal activity. The results showed that the HJC protein expressed from E.coli was substantially equivalent to that of a transgenic rice carrying an HJC gene in terms of immune recognition. The refolded HJC protein exhibited high insecticidal activity against Hyphantria cunea. To the end, this protein can be used to substitute extrinsic plant protein in food-safty assessment of genetically modified products with HJC gene.
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Received: 06 February 2012
Published: 25 June 2012
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