Eukaryotic expression of P-selectin functional segment on the membrane of CHO.
- Author:
Xiaofang LOU
1
;
Shuang WANG
;
Weishi DU
;
Rui YU
;
Weiyuan YU
;
Zhiwei SUN
Author Information
1. Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies;
metabolism;
CHO Cells;
Cell Membrane;
metabolism;
Cloning, Molecular;
Cricetinae;
Cricetulus;
Epidermal Growth Factor;
metabolism;
Glycosylphosphatidylinositols;
metabolism;
Lectins;
metabolism;
P-Selectin;
biosynthesis;
genetics;
immunology;
RNA;
metabolism;
Transfection
- From:
Chinese Journal of Biotechnology
2008;24(7):1162-1167
- CountryChina
- Language:Chinese
-
Abstract:
Cell adhesive molecular P-selectin was cloned, expressed and anchored on CHO cell membrane through GPI for selection specific antibodies. Total human platelet RNA was extracted and the functional segment of P-selectin gene was cloned by RT-PCR. The P-selectin functional segment gene was cloned into a eukaryotic expression vector pMCEw2-GPI containing an attenuated neo gene together with a downstream GPI, which was synthesized by overlapping PCR. The recombinant plasmid pMCEw2-GPI-P-selectin was then transfected to CHO(dhfr-) cells and screened with G418. ELISA, western-blot and immunofluorescence were carried out to detect the stability of P-selection expression on cell membrane. These results provided a necessary basis for the following study of selection the antibodies targeting P-selectin.
