Keywords

Jatropha Curcas, Ribosome Inactivating Protein, Expression Vector, Escherichia Coli, Induced Expression

Abstract

Objective: To construct a prokaryotic expression vector for the gene encoding the mature protein of Jatropha curcin and to study its high-efficiency expression conditions. Methods: Design primers based on the cDNA sequence of Jatropha curcas on GenBank, and use PCR to amplify the gene encoding the mature protein of Jatropha curcas toxin from the Jatropha curcas genome; introduce it into the prokaryotic expression vector pQE30, pET32, and It was transformed into E. coli to obtain a recombinant strain. Induce them to produce recombinant proteins under conditions such as different inducers, different temperatures, and different induction times and conduct detection and comparison. Results: The gene encoding the mature protein of Jatropha curcas was amplified from the Jatropha curcas genome using PCR method. The prokaryotic expression vectors pQER and pETR were successfully constructed, and the recombinant strains PRM and PRB were obtained. When induced by different inducers, different temperatures, and different induction times, PRB did not produce recombinant protein, while PRM could express curcin in the form of inclusion bodies. Conclusion: The recombinant bacterial PRM expresses curcin protein with the highest efficiency when induced for 6 hours, 28¡ãC, and IPTG 0ª±5mmol¡¤L-1.

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Original research done by Luo Mengjun 1,2, Chen Fang 2, Yan Quan 2, Liu Weixin 1*

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