含绿色荧光蛋白基因与人疱疹病毒8型K12基因重组真核表达载体的构建  

Construction of Eukaryotic Expression Vector Containing the HHV-8 K12 and Green Fluoresent Protein Gene

作　　者：朱建中[1] 卢春[1] 

机构地区：[1]南京医科大学微生物与免疫学教研室,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2002年第6期453-454,共2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)

摘　　要：目的:构建含绿色荧光蛋白(GFP)基因与人疱疹病毒8型K12基因的重组真核表达载体。方法:将人疱疹病毒8型(HHV-8)K12基因插入到真核表达质粒pCR3.1的EcoRI和XhoI位点,构建重组质粒pCR-K12;PCR扩增GFP基因,将GFP基因插入到重组质粒pCR-K12中K12上游的KpnI和EcoRI位点中,获得重组表达质粒pCR-GFP-K12。结果:重组表达质粒pCR-GFP-K12的酶切鉴定符合预期结果,此重组质粒中的K12与GFP融合表达并受上游启动子pCMV控制。结论:重组表达质粒pCR-GFP-K12已构建成功,为研究K12的生物学活性及其作用机制打下基础。Objective: To construct eukaryotic expression vector containing the HHV 8 K12 gene and green fluoresent protein gene.Methods: The K12 gene of HHV 8 digested with EcoRI and XhoI was inserted into MCS sites of eukaryotic expression vector pCR3.1 to construct pCR K12; Green fluoresent protein (GFP) gene was amplified by polymerase chain reaction (PCR) from pGFPuv as template, digested with KpnI and EcoRI and then inserted into upstream of K12 of pCR K12 to construct pCR GFP K12. Results: Restriction endonucleases digestion of pCR GFP K12 was the same as expected, in which K12 was fused with GFP and controlled by promoter CMV. Conclusion: Construction of pCR GFP K12 was successful, which would benefit study on biology activity and action mechanism of HHV 8 K12.

关 键 词：绿色荧光蛋白 人疱疹病毒8型 K12基因 重组真核表达载体 构建 

分 类 号：R373.11[医药卫生—病原生物学] Q782[医药卫生—基础医学]