The Construction and Expression of Eukaryotic Expression Vector Harboring Homo Sapiens Integrin beta 3 ORF Gene Segment

To study the cellular entry of Hantavirus, we constructed dukaryotic expression vectors harboring Homo sapiens integrin β3 gene. The 2.3kb ORF region of human integrin β3 subunit cDNA was amplified from the plasmid containing cDNA of human integriri β3 by PCR. Then the PCR products with the blunt-ends were directly cloned into eukaryotic ecpression vector pcDNA 3.1/V5-His TOPO by TA Cloning. It shows that the integrin gene was inserted into the plasmid vector by restriction enzyme analysis and PCR, and the sequence is identical with the reported human integrin β3 ORF gene, then the recombinant expression plasmids were transfected into CHO cells in vitro. The transient expression products were analyzed by IFA. The results above suggested that the eukaryotic expression vectors, pcDNA3.1-β3, has been successfully constructed and the expressed protein could be detected distinctly and peculiarly. It lies a basis for the further study of hantavirus pathogenicity and biological characteristics