Construction of a recombinant eukaryotic expression plasmid containing human PDLIM2 gene and its biological activity.
Plasmid
; 66(2): 106-11, 2011 Jul.
Article
en En
| MEDLINE
| ID: mdl-21784097
In order to obtain a full-length expression plasmid for human PDLIM2 gene, fragment amplification was used to clone its full-length coding sequence (CDS) region. The amplified PCR product was then digested and inserted into the pMD 18-T vector and subcloned into the pIRES2-EGFP plasmid to form the pIRES2-EGFP-PDLIM2 eukaryotic expression vector. After it was transfected to the bladder cancer cell line, BIU-87, the biological activities of high expression were verified by RT-PCR and Western blotting. Meanwhile the mRNA and protein expressions of p65 were detected. Finally we analyzed the effect of overexpressed PDLIM2 on BIU-87 cell proliferation. In conclusion, a recombinant eukaryotic expression vector pIRES2-EGFP-PDLIM2 containing the complete CDS region of PDLIM2 was successfully constructed. PDLIM2 negatively regulated p65 expression and inhibited BIU-87 cell proliferation. We laid the foundations for further research into the function of the PDLIM2 gene in bladder cancer.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Plásmidos
/
Proteínas con Dominio LIM
/
Proteínas de Microfilamentos
Límite:
Humans
Idioma:
En
Revista:
Plasmid
Año:
2011
Tipo del documento:
Article
Pais de publicación:
Estados Unidos