Construction and Virulence Analysis of MocR Transcription Regulator <em>SSU0562</em> Gene Knock-out Mutant in <em>Streptococcus suis</em> Serotype2

doi:10.13523/j.cb.20150702

China Biotechnology

2015, Vol. 35

Issue (7): 8-14 DOI: 10.13523/j.cb.20150702

Construction and Virulence Analysis of MocR Transcription Regulator SSU0562 Gene Knock-out Mutant in Streptococcus suis Serotype2

LI Juan^1,2, LIU Li-na², HU Dan¹, ZHU XU-hui¹, GONG Xiu-fang¹, ZHAO Lin¹, ZHONG Jing-hao¹, PAN Xiu-zhen¹, WANG Chang-jun¹

1. Institute of Military Medical Sciences, Nanjing Command, Nanjing 210002, China;
2. Guiyang Medical University, School of Biology & Engineering, Guiyang 550004, China

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Abstract

Objective:After construction of the knock-out mutant of MocR family transcription regulator SSU0562 gene in Streptococcus suis serotype 2, the virulence and biological characteristics of the mutant strain was analysised. Methods:Based on the principle of homologous recombination,recombinant gene knock-out vector was constructed consisting of Spc^r cassette with the flanking homology regions to the target gene and the isogenic SSU0562-difficient mutant was screened by allelic replacement. The basic biological characteristics between the mutant Δ0562 and the wild type strain 05ZYH33, and the virulence in murine model were compared. Results:PCR analysis and gene sequencing results confirmed the SSU0562 gene was completely replaced by Spc^r cassette, showing the gene knock-out mutant strain Δ0562 was successfully constructed. Reverse transcription PCR (RT-PCR) confirmed the absence of SSU0562 in the Δ0562 strain at the transcriptional level. Study of biological characteristics and the virulence showed that there were no significant differences between Δ0562 and 05ZYH33 in terms of hemolytic activity, growth kinetics and pathogenicity. The chain ability of Δ0562 was significantly weaker than 05ZYH33. Conclusion: SSU0562 gene knock-out did not change the virulence of Streptococcus suis serotype 2 05ZYH33, showing that SSU0562 is not the virulence decision factor of Streptococcus suis. The SSU0562 gene was probably involved in the regulation of the chain ability in Streptococcus suis.

Key words： Streptococcus suis serotype 2 MocR SSU0562 Gene knock-out virulence

Received: 25 December 2014 Published: 25 July 2015

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LI Juan, LIU Li-na, HU Dan, ZHU XU-hui, GONG Xiu-fang, ZHAO Lin, ZHONG Jing-hao, PAN Xiu-zhen, WANG Chang-jun. Construction and Virulence Analysis of MocR Transcription Regulator SSU0562 Gene Knock-out Mutant in Streptococcus suis Serotype2. China Biotechnology, 2015, 35(7): 8-14.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150702 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I7/8

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