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Chinese Journal of Biotechnology ; (12): 1162-1167, 2008.
Artigo em Chinês | WPRIM | ID: wpr-275409

RESUMO

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.


Assuntos
Animais , Cricetinae , Anticorpos , Metabolismo , Células CHO , Membrana Celular , Metabolismo , Clonagem Molecular , Cricetulus , Fator de Crescimento Epidérmico , Metabolismo , Glicosilfosfatidilinositóis , Metabolismo , Lectinas , Metabolismo , Selectina-P , Genética , Alergia e Imunologia , RNA , Metabolismo , Transfecção
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