熊源粪肠球菌EfaA基因的原核表达

doi:10.16431/j.cnki.1671-7236.2018.01.009

Abstract

Abstract:

This study was aimed to construct and express the prokaryotic expression plasmid of EfaA gene. One pair of PCR primers was designed according to the EfaA gene sequence from GenBank (accession No.:U03756). EfaA was used as target gene to clone and analyze by biological information. EfaA gene was sub-cloned into pGEX-4T-1, PGEX-4T-EfaA prokaryotic expression plasmid was constructed, and the plasmid was transformed into E.coli BL21(DE3), which was induced by IPTG. The best induction time was screened and biological characteristics of the protein was analyzed. The results showed that 873 bp fragment of Enterococcus faecalis EfaA gene was obtained successfully. Induced expression was operated by IPTG and 59 ku EfaA recombinant protein was obtained. The best way of this react was 6 h in 37℃ by IPTG (0.5 mmol/L). The Western blotting result showed good reactogenicity. The prokaryotic expression plasmid pGEX-4T-EfaA was successfully constructed and expressed in E.coli BL21 (DE3).

Key words: Black bear; Enterococcus faecalis; endocarditis antigen; prokaryotic expression

CLC Number:

S852.61⁺1

CHU Xitong, QIN Xiaodong, LU Cheng, SUN Fuliang. Prokaryotic Expression of EfaA Gene of Enterococcus faecium from Bear[J]. , 2018, 45(1): 71-76.

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Prokaryotic Expression of EfaA Gene of Enterococcus faecium from Bear

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