人CETP cDNA的克隆与真核表达载体的构建及鉴定  

cDNA of Human Cholesteryl Ester Transfer Protein Cloning and Construction and Identification of Its Eukaryotic Expression Vector

作　　者：任春秀[1] 佳元[1] 何一平[1] 刘惠荣[1] 

机构地区：[1]内蒙古农业大学生物工程学院,内蒙古呼和浩特010018

出　　处：《畜牧与饲料科学》2008年第4期47-48,52,共3页Animal Husbandry and Feed Science

基　　金：内蒙古农业大学博士科研启动基金项目(BJ05-34)。

摘　　要：通过克隆人CETP cDNA ORF序列,构建人CETP真核表达载体,为进一步研究人CETP的结构与功能奠定基础。该试验提取人肝组织的总RNA并纯化mRNA,用试剂盒将mRNA反转录合成cDNA-链,设计引物,以合成的cDNA链为模板PCR扩增CETP cDNA的ORF序列。将扩增得到的约1.5kb片段克隆进入pcDNA3.1/myc-His(-)A质粒载体,最后将重组质粒进行鉴定并测序。结果表明,CETP重组真核表达载体包含完整的CETP cDNA ORF序列,且与已发表的人CETP cDNA ORF序列完全一致。In order to lay a foundation for further study on the structure and function of CETP, the ORF sequence of human CETP eDNA was cloned and the eukaryotie expression vector was constructed. Total RNA was extracted from human liver tissues and mRNA was purified. Using kit, mRNA was synthesized to cDNA strand through reverse transcription. Primers were designed and the ORF sequence of human CETP cDNA was amplified from synthesized cDNA strand by PCR. And then 1.5 kb eDNA amplified fragment was cloned into pcDNA3.1/myc-His （-）A to form recombinant plasmid vector. Finally, the recombinant plasmid was sequenced after identified. The results showed that the recombinant eukaryotic expression vector contained the complete ORF sequence of CETP eDNA and the sequence was in full accord with the ORF sequence of the reported human CETP eDNA. The eukaryotie expression vector of human cholesteryl ester transfer protein was constructed successfully in this study.

关 键 词：胆固醇酯转运蛋白 CDNA 真核表达载体 

分 类 号：Q523[生物学—生物化学]