人蛋白CcDNA基因的克隆及序列分析  

Cloning and Sequencing of Human Protein C cDNA

作　　者：贾莉玮[1] 吕茂民[1] 沈永才[1] 赵保全[1] 张亚东[1] 章金刚[1] 

机构地区：[1]军事医学科学院输血医学研究所,北京100850

出　　处：《生物技术通报》2003年第4期26-28,32,共4页Biotechnology Bulletin

基　　金：军事医学科学院创新基金资助项目

摘　　要：为实现人蛋白CcDNA在哺乳动物细胞中的表达以及研究其生物学特性 ,针对人蛋白CcDNA序列设计引物 ,运用逆转录聚合酶链反应 (RT PCR)从人胎肝总RNA中钓取人蛋白CcDNA ,将其克隆入 pIRESneo载体中 ,通过酶切和PCR鉴定出重组体并进行测序分析。结果表明 ,获得大小为 1386bp的人蛋白CcDNA基因 ,成功构建人蛋白CcDNA载体 pIRES/hPC ,为进一步进行人蛋白CcDNA的表达和活性鉴定奠定了基础。For researching biological activities and expressing in the mammary tissue culture cells, human protein C cDNA gene was amplified from fetal liver total RNA by RT-PCR. The recombinant plasmid pIRES/hPC was constructed by inserting the human protein C cDNA into pIRESneo and transformed into E.coli DH5α. The positive cloning that contains the cDNA was identified by endonuclease digestion and PCR. Sequencing of pIRES/hPC confirmed the obtained cDNA fragment was identical to human protein C sequence published in the GenBank. These have laid the foundation of further research on the expression and activity of human protein C cDNA.

关 键 词：人蛋白C CDNA基因 克隆 序列分析 逆转录聚合酶链反应 

分 类 号：Q785[生物学—分子生物学]