猪链球菌2型分选酶srtC5基因敲除突变菌株的构建及其特性分析  

Construction and function study of the srtC5 gene encoding sortase knock-out mutant of Streptococcus suis Serotype 2

作　　者：祝令伟[1] 田园[1] 贝为成[2] 齐翀[1] 刘军[1] 孙洋[1] 周博[1] 郭学军[1] 冯书章[1] 

机构地区：[1]中国人民解放军军事医学科学院军事兽医研究所,吉林省人兽共患病预防与控制重点实验室,吉林长春130062 [2]华中农业大学农业微生物学国家重点实验室,湖北武汉430070

出　　处：《中国兽医学报》2011年第7期993-997,共5页Chinese Journal of Veterinary Science

基　　金：国家973项目(2006CB504402)

摘　　要：构建猪链球菌2型(Streptococcus suis serotype 2)强毒株ZY458分选酶srtC5基因敲除突变体并分析其特性。首先通过重叠延伸PCR技术扩增ΔsrtC5片段,该片段携带srtC5编码基因及其上下游部分序列,但是中间缺失212bp序列。随后构建重组自杀载体pSET4s::ΔsrtC5,并将其电转化导入ZY458菌株,通过同源重组获得ΔsrtC5突变体。通过体外的细胞黏附试验和体内的家兔攻毒试验证明ΔsrtC5突变体比较野毒株,其细胞黏附能力和对家兔的致病性都显著降低。Based on the principle of homologous recombination,we constructed gene knock-out mutant of srtC5 gene encoding sortase in Streptococcus suis serotype 2 （SS2） virulent strain ZY458. Recombinant gene knock-out vector pSET4s： ： △srtC5 was constructed consisting of flanking homology sequence to the target region,the isogenic 212 bp deletion mutant was screened by allelic replacement. PCR analysis and sequencing confirmed that the coding gene of srtC5 was replaced by △srtC5, the mutant of ZY458 srtC5 gene was successfully constructed. The results of cell ad- herence assay and rabbits infection experiment showed that the srtC5 deletion mutant （△srtC5） attenuated cellular adhesion ability and the full virulence of its wild-type strain （ZY458）.

关 键 词：猪链球菌2型 分选酶srtC5基因 敲除突变 细胞黏附 

分 类 号：S852.61[农业科学—基础兽医学]