Eukaryotic expression of P-selectin functional segment on the membrane of CHO / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1162-1167, 2008.
Article
in Chinese
| WPRIM
| ID: wpr-275409
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
RNA
/
Transfection
/
Cell Membrane
/
Cricetulus
/
CHO Cells
/
Cloning, Molecular
/
Glycosylphosphatidylinositols
/
P-Selectin
/
Allergy and Immunology
/
Epidermal Growth Factor
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2008
Type:
Article
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