人F3-Restin基因的克隆与表达  

Cloning and Expression of Human F3-Restin Gene

作　　者：刘洋[1] 曾昭淳[2] 

机构地区：[1]长江大学医学院,湖北荆州434023 [2]重庆医科大学生物化学与分子生物学教研室,重庆400016

出　　处：《长江大学学报（自科版）（下旬）》2009年第3期11-13,共3页Journal of Yangtze University

摘　　要：目的:克隆人F3-Restin基因,表达有生物学活性的新型融合蛋白F3-Restin。方法:采用亚克隆技术构建融合表达载体PET32a(+)-F3-Restin,原核诱导表达,经Ni2+-NTA琼脂糖树脂亲和层析纯化,透析去盐。结果:在20℃、10h的条件下IPTG原核诱导表达、纯化,得到分子量约为40kD的高纯度的可溶性蛋白F3-Restin。结论:成功地克隆了人F3-Restin基因,构建融合表达载体PET32a(+)-F3-Restin,并表达出新型融合蛋白F3-Restin,为进一步研究F3-Restin蛋白抗肿瘤血管生长的活性及作用机理奠定了良好的基础。Objective:To clone the DNA sequence coding the F3-Restin,and generate the fused protein F3-Restin.Methods:Prokaryotic expression vecter PET32a(+)-F3-Restin was constructed by subcloning technique,then the fusion protein of F3-Restin was expressed in E.coli and purified using Ni2+NTA agarose resin.Result:The expression of the fusion protein was at the highest level 10h in 20℃ induced by IPTG.After purification,the highly purified fusion protein(Mr 40000) was dissoluble.Conclusion:The F3-Restin prokaryotic expression vecter is successfully constructed,and the target protein is expressed and highly purified.A good foundation is established for the research of F3-Restin in future.

关 键 词：F3-Restin基因 克隆 表达 纯化 

分 类 号：R382.2[医药卫生—医学寄生虫学] R978[医药卫生—基础医学]