欧文氏弧菌毒力蛋白PirAB的原核表达与纯化

对虾肝胰腺坏死病的爆发造成了对虾养殖产业的严重亏损。从上海地区凡纳滨对虾中分离出1株欧文氏弧菌(Vibrio owensii SH-14),该菌株可导致凡纳滨对虾死亡,且死虾出现对虾肝胰腺坏死病的典型症状。经PCR扩增欧文氏弧菌毒力蛋白PirA与PirB对应基因序列,并将其连接到表达质粒pET-21b(pirA)和pGEX-4t-1(pirB)。通过优化诱导表达和亲和层析纯化条件,最终获得大量高纯度的目的蛋白。经冷冻干燥保存为后期抗体合成以及进一步毒力效应研究提供参考。

Prokaryotic Expression and Purification of Virulent Proteins PirAB from Vibrio owensii

The fulmination of acute hepatopancreatic necrosis disease (AHPND) creates severe loss to shrimp farming industry. An AHPND causing strain Vibrio owensii SH-14 was isolated from white prawn (Litopenaeus vannamei) in Shanghai and confirmed to create the death of the shrimp and the shrimp appeared typical symptoms of AHPND. In this study, both pirA and pirB corresponding genes of virulent proteins in the V. owensii SH-14 were amplified by PCR and linked to plasmid vectors of pET-21b and pGEX-4t-1 respectively, and expressed in E. coli. Finally a large scale of the PirAB objective proteins were highly purified with affinity chromatography and frozen dried and conserved for later systhesis of antibody, and laid a foundation for further study on the virulent effect.