SLPI基因真核表达质粒的构建和鉴定

摘要

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.%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peDNA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.