[Construction of human integrin beta3 subunit eukaryotic expression vector and efficient surface expression of integrin alphaIIbbeta3].

ABSTRACT

AIM: To establish efficient surface expression of Homo sapiens integrin alphaIIbbeta3 in beta3-integrin-deficient and HV-insusceptible CHO cells, so as to lay the foundation for the further study of cellular entry of hantavirus mediated by beta3 integrins. METHODS: Eukaryotic expression vector pcDNA3.1-beta3 harboring ORF region of human integrin beta3 subunit cDNA was constructed, then pcDNA3.1-beta3 and eukaryotic expression vector pBJ1-alphaIIb containing human integrin alphaIIb subunit cDNA were transfected into CHO cells alone or together. The expression of exogenous genes were analyzed by indirect immunofluorescence assay (IFA). RESULTS: The eukaryotic expression vector pcDNA3.1-beta3 was constructed successfully. IFA examination showed that surface expression of integrin alphaIIb beta3 was highly effective in cotransfection group, while surface expression was weak on CHO cells transfected with pcDNA3.1-beta3 alone,and no effective surface expression was found in pBJ1-alphaIIb-transfected group. CONCLUSION: Efficient surface expression of integrin alphaIIb beta3 requires expression of both subunits.