新型环形病毒AGVAGV2衣壳蛋白VPVP1的原核表达

Prokaryotic Expression of VP1 Coat Protein of a Novel Avian Gyrovirus 2

为建立AGV2血清学检测方法,利用p GEX-6P-1载体,对AGV2的衣壳蛋白VP1基因进行分段GST融合表达,成功构建了4个VP1分段GST融合表达载体,分别命名为VP1_1-462、VP1_463-924、VP1_925-1383以及VP1_735-1089。经IPTG诱导、SDS-PAGE以及Western blot分析发现,VP1_1-462以及VP1_925-1383的表达产物主要以包涵体形式表达,而VP1_463-924以及VP1_735-1089的表达产物具有可溶性,且VP1_925-1383的表达量最高。这些AGV2 VP1分段GST融合表达产物的获得为建立用于AGV2流行病学调查的血清学检测方法奠定理论基础。

PCR is the only method for detection of the novel avian gyrovirus 2（AGV2）so far,and there is no report for detection of AGV2 by serological approach. To establish the serological method for detection of AGV2,the VP1 coat protein of AGV2 was divided into four parts and expressed as GST fusion proteins in E.coli. And four fusion expression vectors were generated,and named as VP1_1-462,VP1_463-924,VP1_925-1383 and VP1_735-1089.SDS-PAGE and Western blot analysis revealed that the expression products of VP1_1-462 and VP1_925-1383 were mainly located in the pellet as inclusion body after IPTG induction, whereas those of VP1_463-924 and VP1_735-1089 were found both in the pellet and supernatant. Among the four expression products,the expression level of VP1_925-1383 was the highest. Preparation of these GST fusion proteins of AGV2 VP1 provided materials for further developing serological approaches for AGV2 epidemiological surveillance.