SLPI基因真核表达质粒的构建和鉴定  

Construction and identification of eukaryotic expression plasmids containing SLPI gene

作　　者：朱亮华 金蕊 陆超 周国平 胡毓华 

机构地区：[1]南京医科大学第一附属医院儿科,江苏省210029

出　　处：《江苏医药》2012年第11期1247-1249,共3页Jiangsu Medical Journal

基　　金：江苏省卫生厅医学科技发展基金(H201022)

摘　　要：目的构建并鉴定含分泌型白细胞蛋白酶抑制蛋白(SLPI)基因的重组真核表达质粒。方法以HeLa细胞的总RNA为模板,用RT-PCR方法扩增出SLPI基因cDNA,将产物克隆进真核载体pcDNA3.1(+)内,构建含SLPI基因的重组真核表达质粒。将pcDNA-SLPI重组质粒和空载体pcDNA分别转染HeLa细胞,Western blot检测SLPI蛋白表达。结果核酸序列分析及双酶切鉴定SLPI已成功插入pcDNA3.1(+)载体中,转染pcDNA-SLPI的HeLa细胞中检测到高表达的SLPI蛋白。结论成功构建了含SLPI基因的重组真核表达质粒。Objective To construct and identify recombinant eukaryotic expression plasmids containing secretary leukocyte protease inhibitor（SLPI） gene. Methods The cDNA SLPI gene was amplified by RT-PCR with the total RNA in HeLa cells as a template. The PCR fragments were cloned into pcDNA3.1 （＋） vector to construct recombinant eukaryotic expression plasmids containing SLPI gene. The pcDNA-SLPI recombinant plasmid and pcDNA empty vector were respectively transfected into HeLa cells, and the expression of SLPI protein was detected by Western blot. Results The DNA sequence and double restrictive endonuclease analysis showed that SLPI was successfully inserted into pcDNA3.1（＋） vector. The high expression of SLPI protein was detected in HeLa cells transfected with pcDNA-SLPI plasmid by Western blot. Conclusion Recombinant eukaryotic expression plasmids containing SLPI gene has been successfully constructed.

关 键 词：分泌型白细胞蛋白酶抑制蛋白 真核表达载体 

分 类 号：R722[医药卫生—儿科]