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


CONCLUSIONS
Efficient surface expression of integrin alphaIIb beta3 requires expression of both subunits.
Mou
Dan-Lei
DL
Center of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038,China.
Bai
Xue-Fan
XF
Huang
Chang-Xing
CX
Li
Guang-Yu
GY
Pan
Lei
L
Wang
Ying-Peng
YP
Yang
Wei-Song
WS
Zhang
Hong-Fei
HF
D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D056890:Eukaryota; D006801:Humans; D039661:Integrin beta3 / Q000235:genetics; D019039:Platelet Glycoprotein GPIIb-IIIa Complex
D039661:Integrin beta3; D019039:Platelet Glycoprotein GPIIb-IIIa Complex