真核表达质粒EX-Y2069-M 29的构建及LRP16基因在人子宫内膜癌HEC-1-B细胞中的表达  

Construction of the eukaryotic expression plasmid EXY2069-M29 and analysis of LRP16 expression in human endometrial carcinoma HEC-1-B cells

作　　者：伍宗惠[1] 詹平[1] 邹倩[1] 张宇骄[1] 刘玲[1] 

机构地区：[1]泸州医学院附属医院妇产科,四川泸州646000

出　　处：《中国现代医学杂志》2014年第30期8-13,共6页China Journal of Modern Medicine

基　　金：四川省泸州医学院课题(No:07055)

摘　　要：目的构建LRP16基因真核表达质粒EX-Y2069-M29,检测LRP16基因在人子宫内膜癌HEC-1-B细胞中的表达。方法体外培养人子宫内膜癌HEC-1-B细胞,提取总RNA,应用PCR技术,扩增获得LRP16基因编码序列片段,克隆入真核表达载体EX-Y2069-M29,对重组质粒进行酶切和测序鉴定后,采用脂质体法将LRP16真核表达载体EX-Y2069-M29和EX-NEG-M29空质粒转染细胞,利用逆转录聚合酶链式反应(RT-PCR)和Western blot法分析LRP16基因的表达情况。结果经LRP16基因真核表达质粒EX-Y2069-M29转染的HEC-1-B细胞中检测到LRP16基因的表达。结论构建的LRP16基因重组真核表达质粒EX-Y2069-M29转染HEC-1-B细胞后,可在HEC-1-B细胞中稳定表达,为进一步研究LRP16基因奠定了基础。【Objective】To construct the eukaryotic expression plasmid EX-Y2069-M29 and study the LRP16 gene expression in human endometrial carcinoma HEC-1-B cells. 【Methods】The coding sequence of LRP16 was generated by PCR using total RNA extracted from the HEC-1-B cells. LRP16 gene was cloned into the eukaryotic expression vector EX-Y2069-M29. After restriction enzyme analysis and sequence identification, the recombinant plasmid was transfected into HEC-1-B cells with liposome mediated method. The expression of LRP16 gene was detected by RT-PCR and Western blot. 【Results】The expression of LRP16 gene was observed in HEC-1-B cells which were transfected by recombinant plasmids.【Conclusions】Recombinant plasmid EX-Y2069-M29 constructed with LRP16 gene was expressed stably in HEC-1-B cells, which make the further study of the foundation for LRP16.

关 键 词：LRP16基因 真核表达质粒 HEC-1-B细胞 

分 类 号：R737.33[医药卫生—肿瘤]