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Cloning and expression of HSP27 gene / 第三军医大学学报
Article en Zh | WPRIM | ID: wpr-556533
Biblioteca responsable: WPRO
ABSTRACT
Objective To construct HSP27 eukaryotic expression plasmids. Methods Full-length HSP27 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from breast cancer cell line MCS-7 and cloned into eukaryotic expression vector pAAV-MCS. After the recombinant plasmids transfected into NIH3T3 cells, the expression of HSP27 protein in the host cells was characterized by ECL Western blotting. Results Full-length HSP27 gene was amplified by RT-PCR correctly. The correct cloning of HSP27 gene in pAAV-MCS was confirmed by restriction enzyme digestion and sequencing. ECL Western blotting results indicated that the target gene could express in the mammalian cell line NIH3T3. Conclusion Recombinant plasmid HSP27/pAAV-MCS had been cloned successfully, which would provide the foundation for investigating the role and the mechanism of HSP27 in the ischemia precondition of kidney.
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Texto completo: 1 Base de datos: WPRIM Idioma: Zh Revista: Journal of Third Military Medical University Año: 2003 Tipo del documento: Article
Texto completo: 1 Base de datos: WPRIM Idioma: Zh Revista: Journal of Third Military Medical University Año: 2003 Tipo del documento: Article