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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Construction of a recombinant eukaryotic expression plasmid containing human PDLIM2 gene and its biological activity.
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Construction of a recombinant eukaryotic expression plasmid containing human PDLIM2 gene and its biological activity.

机译：含有人PDLIM2基因的重组真核表达质粒的构建及其生物学活性。

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摘要

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.
机译：为了获得人PDLIM2基因的全长表达质粒，使用片段扩增来克隆其全长编码序列（CDS）区域。然后消化扩增的PCR产物，并插入到pMD 18-T载体中，并亚克隆到pIRES2-EGFP质粒中以形成pIRES2-EGFP-PDLIM2真核表达载体。将其转染至膀胱癌细胞系BIU-87后，通过RT-PCR和Western blotting验证了高表达的生物学活性。同时检测p65的mRNA和蛋白表达。最后，我们分析了过表达的PDLIM2对BIU-87细胞增殖的影响。总之，成功构建了包含PDLIM2完整CDS区域的重组真核表达载体pIRES2-EGFP-PDLIM2。 PDLIM2负调控p65表达并抑制BIU-87细胞增殖。我们为进一步研究PDLIM2基因在膀胱癌中的功能奠定了基础。

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