缺氧调控报告载体的构建与鉴定  

Construction and identification of hypoxia regulatory reporter vectors

作　　者：杨洁[1] 李占亭[1] 杜德伟[1] 柴青焕[1] 王丽[1] 张惠中[2] 孙脊峰[1] 

机构地区：[1]第四军医大学唐都医院肾脏内科,陕西西安7100382 [2]第四军医大学唐都医院中心实验室,陕西西安710038

出　　处：《第四军医大学学报》2006年第7期590-592,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30370662)

摘　　要：目的:构建能用来检测缺氧应答启动子转录调控作用的萤火虫荧光素酶报告载体.方法:合成缺氧应答元件(HRE)的寡核苷酸序列及其突变对照,克隆入pCIneo,构建HRE/CMV重组载体.然后采用PCR方法将HRE/CMV启动子定向亚克隆入pGL3Basic.结果:酶切和测序鉴定分析,所克隆的基因产物酶切结果与预期值一致,序列无碱基的突变.结论:成功构建了缺氧应答启动子的荧光素酶报告载体,为进一步研究其缺氧调控作用提供了条件.AIM： To construct the firefly luciferase reporter gene vectors regulated by hypoxia response promoters. METHODS： The oligonucleotides of hypoxia response element （HRE） and a mutation control were synthesized and cloned into pCI-neo vectors to constrct HRF/CMV recombinant vectors. Then HRE/CMV promoters amplified by PCR were subcloned into pGL3-Basic vectors. RESULTS： The integrity reporter vectors were verified by restriction enzyme digestion. Also, the sequences of the vectors were detected and aligned to the expected sequences. CONCLUSION： Luciferase reporter vectors regulated by HRE/ CMV promoters were successfully constructed, which makes it possible to further investigate their potency of hypoxia regulation.

关 键 词：转录 调控 缺氧 反应元件 荧光素酶 报告基因 

分 类 号：R692.5[医药卫生—泌尿科学]