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.