鸭疫里默氏杆菌假定溶血素相关蛋白基因Riean_0415的原核表达及溶血活性检测

PROKARYOTIC EXPRESSION AND DETECTION OF HEMOLYTIC ACTIVITYOF HYPOTHETICAL HEMOLYSIN-RELATED PROTEIN RIEAN_0415 GENE OF RIEMERELLA ANATIPESTIFER

为了分析鸭疫里氏杆菌(Riemerella anatipestifer,RA)假定的含CBS结构域的溶血素相关蛋白(hypothetical hemolysinrelated protein,HRP)的功能,本研究根据血清10型鸭疫里氏杆菌HXb2株Riean_0415基因的核苷酸序列,设计了1对特异性引物,采用PCR扩增了Riean_0415基因,并进行了序列测定与分析。然后将Riean_0415基因克隆到原核表达载体p ET-43.1a,再用亲和层析纯化获得诱导表达的重组蛋白r HRP。溶血活性检测结果表明表达的重组蛋白r HRP具有裂解鸭红细胞的活性。本研究结果为进一步研究r HRP蛋白的功能和鸭疫里默氏杆菌的溶血机制等奠定了基础。

To investigate the biological function of hypothetical hemolysin-related protein （HRP） of Riemerella anatipestifer, the Riean_0415 gene was prokaryotically expressed and hemolytic activity of the recombinant protein was evaluated. The Riean_0415 gene was amplified from serotype 10 strain HXb2 in PCR and cloned into Promega T easy vector. Then, DNA fragments were ligated to the prokaryotic expression vector pET-43.1a. The expression of recombinant protein HRP was induced with IPTG and purified with affinity chromatograph. In red cell lysis test, the recombinant protein HRP had the ability to lyse duck erythrocytes. The result of this study will benefit to further study on biological functions of HRP protein and hemolysis mechanism of Riemerella anatipestifer.