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[Eukaryotic expression of P-selectin functional segment on the membrane of CHO].
- Source :
-
Sheng wu gong cheng xue bao = Chinese journal of biotechnology [Sheng Wu Gong Cheng Xue Bao] 2008 Jul; Vol. 24 (7), pp. 1162-7. - Publication Year :
- 2008
-
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.
- Subjects :
- Animals
Antibodies metabolism
CHO Cells
Cell Membrane metabolism
Cloning, Molecular
Cricetinae
Cricetulus
Epidermal Growth Factor metabolism
Glycosylphosphatidylinositols metabolism
Lectins metabolism
P-Selectin genetics
P-Selectin immunology
RNA metabolism
P-Selectin biosynthesis
Transfection
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1000-3061
- Volume :
- 24
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 18837389