动物医学进展2016，Vol.37Issue(4)：35-39,5.
罗非鱼无乳链球菌 EsxA 基因克隆与原核表达
Clonging and Protokaryon Expression of EsxA Gene of Streptococcus agalactiae in Tilapia
摘要
Abstract
New discovered type Ⅶ secretion system (T7SS)secrets two extracellular proteins including Es-xA coded ESAT6 and EsxB coded CFP-10,which is related to pathogenicity closely.ESAT6 and CFP-10 proteins have diverse functions such as promoting bacteria from phagosome of macrophage,impacting mac-rophage apoptosis and lysis.The two secreted proteins in T7SS of Streptococcus agalactiae form the ho-mologous dimmers,collectively known as ESAT6 protein,may be bacterial adhesion protein showing im-portant for host-cell attachment and invasion in S.agalactiae through bioinformatics analysis.But ESAT6 protein whether has the virulence and its pathogenesis is not clear,therefore it is necessary to study on it. We cloned 294 bp sequence from tilapia Streptococcus agalactiae .And the sequence was cloned into pMD18-T vector,the homology is 99% with tilapia Streptococcus agalactiae EsxA gene.EsxA gene was cloned into pET32a vector.The best solouble expression was under 0.5 mmol/L IPTG,28℃.In addition, the purified protein was used to immunize Balb/c mice to prepare mouse anti-ESAT6 protein antibody.ES-AT6 protein had sastisfied reactogenicity through Western blot test.These results encouraged further re-search for the immunological function of S.agalactiae ESAT6 protein.关键词
Ⅶ型分泌系统/无乳链球菌/EsxA 基因/克隆与表达Key words
type Ⅶ secretion system/Streptococcus agalactiae/EsxA gene/clonging and expression分类
农业科技引用本文复制引用
马艳平,梁志凌,马江耀,刘振兴,郝乐,柯浩..罗非鱼无乳链球菌 EsxA 基因克隆与原核表达[J].动物医学进展,2016,37(4):35-39,5.基金项目
广东省海洋渔业局科技与产业发展专项（A201401C06）；广州市农业科技攻关项目（GZCQC1202FG04002-01）；广州市科技攻关项目 （）
