E2F3基因真核表达载体的构建和表达

Construction and expression of eukaryotic expression vector containing gene E2F3

目的克隆人膀胱肿瘤组织E2F3基因及构建E2F3基因真核表达载体并研究其表达。方法取新鲜膀胱肿瘤组织,提取总RNA,采用逆转录-聚合酶联链反应(RT-PCR)扩增E2F3基因,构建其真核表达载体,测序分析及转染膀胱肿瘤细胞,Western Blot法检测其表达情况。结果成功克隆人膀胱肿瘤组织E2F3基因、构建E2F3基因真核表达载体,转染后成功表达。结论人膀胱肿瘤组织E2F3基因克隆及其真核表达载体构建成功,可用于肿瘤生物学的研究。

[Objective] To clone human bladder tumor E2F3 gene, construct its eukaryotic expression vector and expression in the BIU-87 cell. [Methods] E2F3 gene was obtained from bladder tumor by RT-PCR. Molecular cloning technique was used to construct this kind of eukaryotic expression vector, pEGFP-E2F3 was identified by sequencing. Using liposome-mediated transfection, the eukaryotic expression vector pEGFP-E2F3 was transfected into BIU-87 cell. [Results] The eukaryotic expression vector pEGFP-E2F3 was constructed correctly and was expressed in BIU-87 cell. [Conclusion] These results suggested that this new kinds of eukaryotlc expression vector could serve as a new tool and methods for neoplasm etiology study and gene therapy.