Construction of Prokaryotic Expression Vector of PepA Protein of Vibrio alginolyticus and Identification of Its Acetylation

doi:10.13560/j.cnki.biotech.bull.1985.2020-0444

Abstract

Abstract:

The purpose of this study is to construct a prokaryotic expression vector of PepA protein in Vibrio alginolyticus HY9901,to optimize its expression conditions,and to analyze whether or not it is regulated by protein acetylation. Firstly,the pepA gene was cloned via PCR,and an expression vector pET-28a-PepA was constructed and transferred it into Escherichia coli BL21（DE3）.Then,the protein expression and acetylation were analyzed by SDS-PAGE and Western blot. The recombinant expression strain correctly expressed the recombinant protein（60.7 kD）,and its optimal expression conditions was,induced for 5 h at 37℃ by IPTG with a volume fraction of 0.1%. Western blot results showed that PepA protein was acetylated protein,and there was no deacetylation in vitro.

Key words: Vibrio alginolyticus, PepA protein, optimal expression condition, acetylation

XU Zhou, FAN Chen-long, DING Yu. Construction of Prokaryotic Expression Vector of PepA Protein of Vibrio alginolyticus and Identification of Its Acetylation[J]. Biotechnology Bulletin, 2020, 36(12): 75-81.

Figures/Tables 8

References 28

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