人WWOX基因的克隆及其真核表达载体的构建

目的:克隆人WWOX基因并构建其真核表达载体。方法:从人正常卵巢组织中提取总RNA,经逆转录聚合酶链反应扩增人WWOX基因,并构建其真核表达载体,经测序鉴定后转染人卵巢癌细胞系A2780。结果:成功克隆了人WWOX基因,与Genbank提供的序列(NM-016373)对比显示完全一致。真核表达载体pCMV-WWOX转染人卵巢癌细胞系A2780后,癌细胞的WWOXmRNA的表达水平显著增高(P<0.01)。结论:从人正常卵巢组织中成功克隆了人WWOX基因,为进一步研究WWOX在卵巢癌发生和进展中的作用及其靶向基因治疗奠定了实验基础。

Cloning of human WWOX gene and construction of its eukaryotic expression vector

Objective:To clone human WWOX gene and construct its eukaryotic expression vector.Methods:The total WWOX RNA was abstracted from human normal ovarian tissues,then human WWOX gene was amplified by RT-PCR,its eukaryotic expression vector was constructed,human ovarian cancer cell line A2780 was transfected after gene sequencing.Results:The human WWOX gene was successfully cloned and it was identical to the reported WWOX sequence(NM-016373) provided by Genbank.After the eukaryotic expression vector pCMV-WWOX was transfected into ovarian cancer cell line A2780,the expression level of WWOX mRNA increased significantly(P<0.01).Conclusion:The human WWOX gene is successfully cloned from human normal ovarian tissues,which provides a experimental basis for investigating the effect of WWOX in the occurrence and development of ovarian cancer and targeted gene therapy.