pcDNA3.1(+)gpc-5真核表达载体的构建及鉴定

目的：构建pcDNA3.1（＋）磷脂酞肌醇蛋白5（gpc-5）基因真核表达载体。方法：根据gpc-5基因序列设计一对引物,利用RT-PCR法扩增出gpc-5基因的编码区（CDS）,连接在PGM-T载体上,并进行pcDNA3.1（＋）gpc-5真核表达载体的构建、酶切鉴定以及测序。结果：gpc-5基因的体外扩增目的片段为2 083 bp。所构建的重组体pcDNA3.1（＋）gpc-5经双酶切鉴定,与预期片段大小一致,测序结果与NCB I收录gpc-5基因的CDS序列一致。结论：成功构建了真核表达载体pcDNA3.1（＋）gpc-5,为研究gpc-5基因在原发性肺癌的发生发展中的作用奠定了基础。

Construction and Identification of Eukaryotic Expressing Vector of pcDNA3.1(+)gpc-5

Objective： To construct the recombinant eukaryotic expression vector of pcDNA3.1（＋）gpc-5.Methods： A couple of primers were designed for RT-PCR according to the known sequence of gpc-5 gene.The coding region of gpc-5 gene was amplified by PCR,and PCR product was cloned into PGM-T vector.Then the eukaryotic expression vector of pcDNA3.1（＋）gpc-5 was constructed,and was identified by restricting enzyme digestion analysis and DNA sequencing.Results： The size of amplified gpc-5 gene was 2083bp.Restriction enzyme digestion and DNA sequencing confirmed that recombinant eukaryotic expression vector of pcDNA3.1（＋）gpc-5 had been constructed successfully.The harvested CDS sequence of gpc-5 gene was identical with that registered in the NCBI database.Conclusion： The eukaryotic expression vector of pcDNA3.1（＋）gpc-5 has been constructed successfully,which may lay the foundation for further studies on the roles of gpc-5 gene in the development of lung cancer.