含mdr1启动子的荧光素酶报告基因质粒的构建

The construction of a luciferase reporter gene vector directed by mdr1 promoter

目的克隆编码mdr1基因的启动子并插入荧光素酶报告基因载体。方法用PCR技术扩增出人类mdr1基因启动子片段,通过亚克隆将启动子分别插入到pGEMT载体和荧光素酶报告基因pGL3enhancer载体中,并确定扩增DNA的序列。结果测序结果显示扩增mdr1启动子序列正确。结论成功克隆了mdr1启动子,为下一步对多耐药性卵巢癌进行靶向基因治疗提供了重要基础。

Objective To clone the promoter of multi-drug resistance 1 gene and insert it into a luciferase reporter gene vector. Methods The promoter of the human mdr1 gene was amplified from human genomic DNA by PCR, and then it was subcloned into pGEM-T and pGL3-enhancer. The plasmid was determinded by DNA sequencing. Results The result of DNA sequencing showed that the sequence of cloned promoter is right. Conclusion the promoter of mdr1 was Cloned successfully,and it will be a important basis for the study of target suicide gene therapy of cisplatin-resistant ovarian cancer.