title: Construction of a recombinant eukaryotic expression plasmid containing human PDLIM2 gene and its biological activity.
abstract: 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.
authors.affiliation: Department of Urology, Anhui Provincial Hospital, Anhui Medical University, Hefei, Anhui, People's Republic of China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xiao
authors.forename: Ling-Wu
authors.initials: LW
authors.lastname: Chen
authors.forename: Jun-Xing
authors.initials: JX
authors.lastname: Chen
authors.forename: Huan-Yi
authors.initials: HY
authors.lastname: Lin
authors.forename: Bin
authors.initials: B
authors.lastname: Huang
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003001:Cloning, Molecular; D005822:Genetic Vectors; D006801:Humans; D060588:LIM Domain Proteins / Q000235:genetics* / Q000378:metabolism*; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000473:pathology
chemical_list: D060588:LIM Domain Proteins; D008840:Microfilament Proteins; C502929:PDLIM2 protein, human; D011994:Recombinant Proteins; D051996:Transcription Factor RelA