CAJ | 학술논문

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。
목적연구잡파육류상관포진병독( KSHV) ORFK12기인재대장간균적표체급표체적단백재보통인군중검측KSHV적감염정황。방법설계일대특이성인물，이BCBL-1세포총DNA위모판，채용PCR방법확증KSHV ORFK12기인。구건함목적기인적중조질립명명위PQE-80L-K12화유도단백표체。채용ELISA법화Western blot법사사림상수집적혈청표본중감염KSHV적정황。결과이병기-β-D-류대필남반유당감(IPTG)유도후적PQE-80L-K12중조균경십이완기류산납-취병희선알응효전영( SDS-PAGE )현시유일개약10 ku적융합표체단백위 ORFK12단백。채용 ELISA법화Western blot법검측현시ORFK12단백능여KSHV양성혈청반응。결론KSHV ORFK12기인재대장간균중표체，표체적ORFK12단백재실험실검측감염KSHV유일정적보조작용。
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.