猪链球菌4型GalR蛋白的生物信息学分析及原核表达

doi:10.16431/j.cnki.1671-7236.2021.02.005

Abstract

Abstract: The aim of this study was to understand the bioinformatics information of GalR protein,a transcription regulator of Streptococcus suis serotype 4 (SS4),and to express it in prokaryotic cells.The sequence homology and functional domain of GalR protein of SS4 SH1510 strain were searched by bioinformatics analysis website,and the signal peptide,transmembrane domain and isoelectric point were also predicted.At the same time,the GalR gene of SS4 SH1510 strain was amplified,prokaryotic expressed and purified.SDS-PAGE was used to analyze the solubility of the recombinant protein,Western blotting was used to identify the reactivity of the recombinant protein.The amino acid sequence alignment showed that it had 57%,56%,55%,55%,52%,52% and 49% homology with the GalR family proteins of Streptococcus mutans,Streptococcus agalactiae,Pseudomonas aeruginosa,Streptococcus equi subsp.zooepidemicus,Streptococcus pneumoniae, Streptococcus oralis and Listeria,respectively.Domain search showed that the GalR protein had a small DNA binding domain with a helix-turn-helix (HTH) motif at the N-terminus,and a ligand-regulated binding of type Ⅰ periplasmic binding protein folding at the C-terminus.In the middle of these two functional domains,there was a linker of about 18 amino acids.GalR protein had no signal peptide and transmembrane region,and its isoelectric point was 5.10.SDS-PAGE analysis showed that GalR protein was mostly expressed in the supernatant and the molecular weight was 56 ku.Western blotting identification showed that His monoclonal antibody and crude SS4 polyclonal rabbit serum could specifically recognize soluble GalR protein.A comprehensive bioinformatics analysis of the GalR gene was performed and the GalR protein was successfully expressed and purified in this study,which laid a foundation for further study of the role of GalR protein in SS4.

Key words: Streptococcus suis serotype 4 (SS4); transcriptional regulator GalR; bioinformatics; prokaryotic expression

CLC Number:

S852.61

SUN Ke, ZHU Haodan, ZHOU Junming, WANG Dandan, YU Zhengyu, LYU Lixin, HE Kongwang, LI Bin, NI Yanxiu. Bioinformatics Analysis and Prokaryotic Expression of GalR Protein of Streptococcus suis Serotype 4[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(2): 443-449.

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Bioinformatics Analysis and Prokaryotic Expression of GalR Protein of Streptococcus suis Serotype 4

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