汉滩病毒核蛋白羧基端多肽原核表达载体的构建及表达

本文为建立分型检测方法，探讨了汉滩病毒(Hantaan virus，HTNV)核蛋白羧基端多肽的抗原性。首先，分别构建编码HTNV核蛋白及其羧基端多肽的原核表达载体pRSETA—S、pRSETA—S—C；然后，将其转化入表达菌BL21(DE3)pLySs诱导表达，采用SDS—PAGE、Westem—blot进行鉴定。我们成功构建了pRSET A—S及pRSETA—S—C原核表达载体。SDS-PAGE显示目的蛋白大量表达，呈不溶状态，Westem—blot显现目的蛋白具有良好的抗原性。为大量制备分型用核蛋白多肽抗原创造了条件。

Construction of Prokaryotic Expression Vectors Harboring HV S and S Partial Gene Segments and Identification of Expressed Nucleocapsid Protein

To study the antigenicity of prokaryotic expressed products of Hantavirus(HV) S partial gene(502-1326bp) and obtain data for serotyping antigen, the prokaryotic expression vectors pRSET A-S, pRSET A-S-C were constructed respectively. The recombinant expression plasmids were transformed into BL21(DE3)pLySs. The expression product were analyzed by SDS-PAGE and Western-blot. The results showed that the proteins were expressed at high level respectively and could be recognized by patients ' (infected by HTNV) sera distinctly and peculiarly. The truncated NP (aa155 to 429) could be considered as serotyping antigen.