布鲁氏菌S19毒力基因BvfA的原核表达及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2021.08.007

Abstract

Abstract: The objective of this study was to clone and express virulence gene BvfA of Brucella (B. abortus) S19, and analyzed the bioinformatics of its expressed protein.In the tests, chromosome 1(accession No.:CCP030751.1)of B.arbortus S19 was downloaded from GenBank.According to the reported position of BvfA gene and molecular weight of BvfA protein in B.arbortus S19, the Open Read Frame (ORF) of BvfA proteins in NCBI's ORF Finder was predicted.The obtained BvfA gene was ligated into prokaryotic expression vector pET-32a(+) to construct the recombinant plasmid pET-32a(+)-BvfA, and expressed in Escherichia coli BL21(Ril).The expressed products were analysed by SDS-PAGE and Western blotting, the structure of BvfA protein was predicted by bioinformatics methods.The results showed that the full-length ORF of BvfA gene was 336 bp, and the expressed fusion BvfA protein was about 11 ku.Bioinformatics analysis showed that BvfA was an unstable hydrophilic protein with no transmembrane structure and its signal peptide region was 1-28 amino acid, BvfA protein mainly located in the extracellular protein (55.6%).There were 12 potential phosphorylation sites, and no O-glycosylation sites.BvfA protein secondary structure was mainly composed of alpha helix and random coil.The results of this study provided some references for the further study of the pathogenic mechanism of BvfA protein in Brucella and search for new therapeutic targets for brucellosis.

Key words: Brucella; BvfA; prokaryotic expression; bioinformatics analysis

CLC Number:

S852.61⁺4

YANG Jiangliu, QIAO Junwei, SONG Qingshan, JIA Fang. Prokaryotic Expression and Bioinformatics Analysis of Virulence Gene BvfA of Brucella S19[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2746-2753.

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Prokaryotic Expression and Bioinformatics Analysis of Virulence Gene BvfA of Brucella S19

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