Cloning of cry2Aa and cry2Ab genes from new isolates of Bacillus thuringiensis and their expression in recombinant Bacillus thuringiensis and Escherichia coli strains |
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| Authors: | Shantanu Kumar V Udayasuriyan |
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| Institution: | (1) Centre of Advanced Studies in Agriculture Microbiology, Tamil Nadu Agricultural University, Coimbatore, 641 003 Tamil Nadu, India;(2) Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore, 641 003 Tamil Nadu, India |
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| Abstract: | Bacillus thuringiensis (Bt) is the major source for transfer of genes to impart insect resistance in transgenic plants. Cry2A proteins of Bt are promising candidates for management of resistance development in insects due to their difference from the currently used Cry1A proteins, in structure and insecticidal mechanism. Two insecticidal crystal protein genes of Bt, viz. cry2Aa and cry2Ab were cloned from new isolates of Bt, 22-4 and 22-11, respectively. Expression of both the genes was studied in an acrystalliferous strain of Bt (4Q7) by fusing the cry2Aa and cry2Ab genes downstream of cry2Aa promoter and orf1 + orf2 sequences. Western blot analysis revealed a low level expression of the cloned cry2Aa and cry2Ab genes in the recombinant Bt strains. High-level expression of cry2Aa and cry2Ab genes was achieved in the recombinant E. coli by cloning the cry2A genes under the control of the T7 promoter. |
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| Keywords: | Bacillus thuringiensis Cry2Aa Cry2Ab E coli T7 promoter |
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