卡波肉瘤相关疱疹病毒ORFK12蛋白初步研究

摘要

目的研究卡波肉瘤相关疱疹病毒( KSHV) ORFK12基因在大肠杆菌的表达及表达的蛋白在普通人群中检测KSHV的感染情况。方法设计一对特异性引物，以BCBL-1细胞总DNA为模板，采用PCR方法扩增KSHV ORFK12基因。构建含目的基因的重组质粒命名为PQE-80L-K12和诱导蛋白表达。采用ELISA法和Western blot法筛查临床收集的血清标本中感染KSHV的情况。结果异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导后的PQE-80L-K12重组菌经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳( SDS-PAGE )显示有一个约10 ku的融合表达蛋白为 ORFK12蛋白。采用 ELISA法和Western blot法检测显示ORFK12蛋白能与KSHV阳性血清反应。结论KSHV ORFK12基因在大肠杆菌中表达，表达的ORFK12蛋白在实验室检测感染KSHV有一定的辅助作用。%Objective To study the expression of ORFK12 gene of KSHV in E. coli and obtain KSHV seropreva-lence in the general population in using the expressed protein. Methods A pair of primers was designed to amplify ORFK12 gene by PCR, using the total DNA of BCBL-1 cells as template. Recombinant plasmid with the target gene was named PQE-80L-K12 and induced to express. In this study, antibodies to one protein of KSHV (ORFK12) were tested by indirect ELISA and Western blot. Results After using IPTG to induce express, PQE-80L-K12 recombinant bacteria had a fusion of 10 Ku protein by SDS-PAGE electrophoresis. 10 specimens were tested. Conclusion It is preliminarily proved that KSHV ORFK12 gene can express in E. coli, and ORFK12 pro-tein infection KSHV plays certain supplementary role in laboratory tests.