人穿孔素真核表达载体的构建及其在COS-7细胞中的表达  

Construction of the eukaryotic expression vector containing the human perforin gene and Its expression in COS-7 cells

作　　者：张春华[1] 李芳秋[1] 缪家文[1] 

机构地区：[1]南京军区南京总医院解放军检验医学研究所,江苏南京210002

出　　处：《医学研究生学报》2008年第2期115-117,共3页Journal of Medical Postgraduates

基　　金：南京市科技发展计划自然科学基金资助项目(批准号:200401070-5)

摘　　要：目的:观察克隆的人穿孔素(pfn)基因在COS-7细胞中的表达情况。方法:用PCR方法获取人pfn全长基因,将所获基因克隆入真核表达载体pcDNA3.1(+),转染COS-7细胞,RT-PCR检测pfn基因在COS-7细胞中的表达。结果:成功获得人pfn全长基因,并构建了真核表达载体。转染COS-7后,检测出pfn基因的表达。结论:人pfn全长基因可以在转染的COS-7细胞中表达。Objective ： To clone human perforin （pfn） full-length DNA, construct the eukaryotic expression vector and observe the expression of the pfn gene in the transfeeted COS-7 cells. Methods： Fulllength DNA ofpfn was obtained by PCR from rPCR2.1/pfn, inserted into the pMD-18T vector and sub-cloned to the pcDNA3.1 （ ＋ ） vector to construct the recombinant eukaryotic expression vector pcDNA3. 1 （ ＋ ）/pfn. The recombinant plasmid was transfected into COS-7 cells and the expression of the pfn gene in the transfected cells was detected by RT-PCR. Results： The pfn full-length DNA was successfully cloned and inserted into the pcDNA3. 1 （ ＋ ） vector. The expression of pfn mRNA in the transfected COS-7 cells was confirmed by RT-PCR. Conclusion ： The full-length human pfn gene can be expressed in transfected COS-7 cells.

关 键 词：穿孔素 真核表达 

分 类 号：Q782[生物学—分子生物学]