Synthesization,Expression and Bioassay of Cry1Ab Insecticidal Gene

doi:10.7668/hbnxb.2014.06.023

Abstract

Abstract: The objective of this study was to modify the coding regions of wild Bacillus thuringiensis Cry1Ab,and then to construct its prokaryotic expression vector and the protein expression in E.coli and insect assay with Asian corn borer.The coding regions of wild Bacillus thuringiensis Cry1Ab was optimized and modified according to plant preferred codons and frequency,and then to synthesize the modified gene (Cry1Ab).Analysis of the DNA sequences revealed that the modified gene was 1 848 bp in length and had 66% percent homology with the native cry1Ab gene,and G+C content was raised from 37.3% to 63.04%.The modified Cry1Ab gene were artificially synthesized and ligated into prokaryotic expression vector pET28b(+) and transformed into E.coli BL21(DE3).SDS-PAGE analysis and about 70 kDa specific fusion protein was produced.Results of insect assay with Asian corn borer showed the expression products of Cry1Ab protein in E.coli had a toxicity to Asian corn borer.Its mortality rate is 93.33% and the growth of the survival larvae were seriously inhibited.We would like to recommend the modified Cry1Ab gene as the candidate for biotechnological crop development.

Key words: Cry1Ab, Synthesis, Prokaryotic expression, Asian corn borer, Bioassay

CLC Number:

YUE Run-qing, TIE Shuang-gui, QI Jian-shuang, HAN Xiao-hua, YAN Shu-feng. Synthesization,Expression and Bioassay of Cry1Ab Insecticidal Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(6): 131-135. doi: 10.7668/hbnxb.2014.06.023.

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