Antigenic Epitope Analysis and Preparation of Polyclonal Antibody of Lepidopteran Pest-resistant Gene cry1C

doi:10.13560/j.cnki.biotech.bull.1985.2018-0200

Abstract

Abstract: In this study,cry1C antigenic epitope region of cry1C was predicted by bioinformatics analysis and designated as Δcry1C. The nucleotide sequence of the Δcry1C was synthesized after codon optimization,and then recombinant expression vector pET28b（+）-Δcry1C was constructed. To prepare cry1C polyclonal antibody,His₆-Δcry1C recombinant protein was purified after induction in Escherichia coli competent cell BL21（DE3）,and then was used to immunize New Zealand white rabbits as antigen. ELISA assay showed that the titer of cry1C polyclonal antibody reached 1：512 000.Western Blot analysis showed that cry1C polyclonal antibody was able to specifically detect the target protein of cry1C in transgenic pest-resistant rice. In summary,cry1C antigenic epitope region was screened by bioinformatics analysis and its polyclonal antibody was prepared successfully. This study provides a basis for further studying Lepidopteran pest-resistant transgenic plants.

Key words: Lepidopteran pest, cry1C, antigenic epitope, codon optimization, polyclonal antibody

JIN Yong-mei, CHEN Mo-jun, LIU Xiao-xiao, LIN Xiu-feng. Antigenic Epitope Analysis and Preparation of Polyclonal Antibody of Lepidopteran Pest-resistant Gene cry1C[J]. Biotechnology Bulletin, 2018, 34(9): 224-229.

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