人疱疹病毒8型K12基因在大肠杆菌中的克隆和表达  

Cloning and Expression of K12 Gene of HHV-8 in E.coli

作　　者：朱建中[1] 卢春[1] 

机构地区：[1]南京医科大学基础医学院病原生物系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2002年第4期274-276,共3页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助项目(30100160);江苏省教育厅科研基金资助项目(99KJB310002);南京医科大学科技发展基金资助项目(NY200001)

摘　　要：目的:将人疱疹病毒8型(HHV-8)K12基因在大肠杆菌中进行融合表达。方法:设计一对PCR引物,在引物的5'端分别引入EcoRI和XhoI酶切位点,用此引物从质粒pCR-K12中扩增出HHV-8的K12基因,PCR产物经双酶切后按正确阅读框连接到原核表达载体pGEX-6p-1上相应酶切位点,并转化BL21感受态细胞。结果:重组菌经异丙基硫代-β-D-半乳糖苷(IPTG)诱导后,菌体SDS-聚丙烯酰胺凝胶电泳(PAGE)显示有一个大小为33ku的融合表达蛋白产生。结论:初步表明K12基因在大肠杆菌中获得正确表达。Objective:To express K12 gene of HHV-8 in E.coli.Methods:A pair PCR primers for HHV-8 K12 gene was designed, in which EcoRI and XhoI sites were introduced at the two 5′ ends of primers respectively. K12 gene was amplified from the pCR-K12 and then inserted into pGEX-6p-1 with proper reading frame. The ligation was transformed into host BL21 cells. Result:SDS-PAGE of the recombinant bacteria showed that a fusion protein of 33 ku was visualized as expected.Conclusion: It suggested that K12 was correctly expressed in E.coli BL21 cells.

关 键 词：人疱疹病毒8型 K12基因 原核表达 

分 类 号：R373.11[医药卫生—病原生物学]