对虾白斑综合征病毒vp28基因的表达及其与血细胞的结合  

Expression and binding activity of vp28 of white spot syndrome virus

作　　者：陈文博[1,2] 谭珍一 刘庆慧[1] 侯林[2] 黄倢[1] 

机构地区：[1]农业部海洋渔业资源可持续利用重点开放实验室中国水产科学研究院黄海水产研究所,青岛266071 [2]辽宁师范大学,大连116029

出　　处：《渔业科学进展》2009年第4期44-49,共6页Progress in Fishery Sciences

基　　金：国家自然科学基金项目(30871942);国家重点基础规划项目(973)(2006CB101801);国家高技术研究发展计划课题(2006AA100312)共同资助

摘　　要：根据GenBank上WSSV囊膜蛋白基因vp28的序列,设计合成引物,PCR扩增得到vp28基因,成功构建重组表达载体pBAD/gⅡIA-VP28并转化大肠杆菌E.coli。用L-阿拉伯糖在37℃诱导重组基因工程菌,表达产物经SDS-PAGE和Western-blot检测,显示有与预期大小31kDa相符合的目的蛋白。荧光显微镜方法分析显示,表达的VP28可与克氏原螯虾血细胞结合。结果表明,在合适的培养条件下,构建的重组表达载体pBAD/gⅡIA-VP28不仅能够表达vp28基因,而且表达的VP28具有很高的抗原结合活性。In this study, a pair of primers was designed by primer design software 5.0 ac- cording to the sequence of vp28 gene of white spot syndrome virus （WSSV） in GenBank. The vp28 DNA fragment was amplified by PCR and cloned into Escherichia coli （E. coli） expression vector pBAD/g Ⅲ A successfully. The pBAD/g Ⅲ A-VP28 was then transformed into E. coli. Af- ter L-arabinose induction at 37℃,the fusion protein with 31 kDa was expressed,which was con- firmed by SDS-PAGE and Western-blot analysis. Binding assay by fluorescence microscopy in vitro showed that VP28 was capable of binding to shrimp hemocytes. Results confirmed that the recombinant vector pBAD/gⅢ A-VP28 can express vp28 gene of WSSV and the VP28 had high antigenic activity.

关 键 词：白斑综合症病毒 VP28 表达 活性分析 

分 类 号：S945.1[农业科学—水产养殖] Q959.223.5[农业科学—水产科学]