对虾白斑综合症病毒结构蛋白VP28的原核表达和性质研究

RESEARCH ON THE PROKARYOTIC EXPRESSION AND CHARACTERISTICS OF VP28,A STRUCTURE PROTEIN OF PENAEUS MONODON WHITE SPOT SYNDROME VIRUS

VP2 8是对虾白斑综合症病毒 (Whitespotsyndromevirus ,WSSV)的一个重要的囊膜蛋白。为了便于研究VP2 8在和宿主细胞相互作用过程中扮演的角色 ,将VP2 8基因克隆到一个原核表达系统 ,对原核表达的VP2 8的特性进行了研究 ,并制备了抗VP2 8的多克隆抗体和单链抗体 ,对原核表达的和天然病毒的VP2 8蛋白免疫原性进行了比较。结果表示 ,原核表达的VP2 8与天然蛋白具有相似的免疫原性。

White Spot Syndrome Virus(WSSV)is an enveloped DNA virus ,which causes 100% cumulative mortality of cultured shrimps in 3 4 days. The virus particles contain five major proteins:VP28、VP26、VP24、VP19 and VP15. The mode of entry and systemic infection of WSSV in shrimp and the functions of these virion proteins are not clear. But more and more attention is paid to the function of VP28,which is likely to play a key role in the initial steps of WSSV infection in shrimps. Towards simplifying to study the role of VP28 in the interaction between WSSV and host cells,the gene of VP28 has been cloned into a prokaryotic expression system,induced and expressed for further researching its characteristics.This prokaryoti expression product could not secrete out of the cell membrane,because it is a membrane protein. Rabbit antiserum against prokaryotic expressed VP28 could recognize both prokaryotic expressed VP28 and native WSSV;Mouse antiserum against native WSSV could recognize both WSSV and prokaryotic expressed VP28. A scFv antibody, designated B7. selected from phage display antibody library against WSSV by panning with prokaryotic expressed VP28,could also recognize both prokaryotic expressed VP28 and native WSSV. These data show that the VP28 produced in a prokaryotic expression system has analogous immunogenic characteristics to its native form,although their protein post modification may differ.