人三突变型低氧诱导因子1α真核表达载体的构建及鉴定  

Construction and identification of pShuttle2 vector of human hypoxia-inducible factor-1α with triple mutants

作　　者：赖艳娴[1] 童锴[1] 谢宜军[1] 王月刚[1] 韦莉莉[1] 吴平生[1] 

机构地区：[1]南方医科大学南方医院心内科,广东广州510515

出　　处：《医学研究生学报》2007年第12期1239-1241,共3页Journal of Medical Postgraduates

基　　金：国家自然科学基金资助项目(批准号:30370587)

摘　　要：目的:为了深入研究人低氧诱导因子1α(HIF-1α)基因对冠心病患者的血管新生作用,构建人三突变型HIF-1α真核表达载体(pShuttle2-HIF-1α-Ala402-Ala564-Ala803)。方法:双酶切pShuttle2-HIF-1α-Ala402-Ala564、pShuttle2-HIF-1α-Ala564-Ala803,胶回收前者酶切片段中300 bp的小片段及后者6300 bp的大片段,并将两者连接,重组成三突变型穿梭质粒pShuttle2-HIF-1α-Ala402-Ala564-Ala803,并进行酶切和PCR鉴定。结果:经酶切鉴定及基因测序证实,重组三突变型穿梭质粒构建成功。结论:成功构建重组人三突变型HIF-1α真核表达载体pShut-tle2-HIF-1α-Ala402-Ala564-Ala803,为进一步促进基因治疗缺血性心脏病的研究奠定基础。Objective ： To construct the pShuttle2 vector of the human hypoxia-inducible factor （HIF）-1α with triple mutants for further study of its effect on therapeutic angiogenesis in patients with coronary heart disease. Methods： pShuttle2-HIF-1α-Ala402-Ala564 and pShuttle2-HIF-1α-Ala564-Ala803 were double-digested for the gel extraction of the fragments, 300bp from the former and 6 300 bp from the latter. The two fragments were connected to form the recombinant shuttle plasmid pShutde2-HIF-1α- Ala402-Ala564-Ala803, and assessment was conducted by digestion and PCR amplification. Results： Restriction endonuclease analysis and PCR amplification confirmed that the recombinant was correctly constructed. Conclusion ： The successful construction of the pShuttle2 vector of human HIF-1α with triple mutants may help prepare the ground for further studies on the gene therapy of coronary atherosclerotic heart diseases.

关 键 词：低氧诱导因子-1Α 基因治疗 血管新生 

分 类 号：Q782[生物学—分子生物学]