小鼠OX2βR突变体表达载体的构建  

Construction of mouse OX2βR mutant expression plasmid

作　　者：潘衍有[1] 王春梅[1] 陈京[1] 白波[1] 

机构地区：[1]济宁医学院泰山学者实验室,山东济宁272067

出　　处：《济宁医学院学报》2013年第5期313-316,319,共5页Journal of Jining Medical University

基　　金：国家自然科学基金项目(编号:81241052;81070961;31271243);济宁市科技局项目(编号:2012jnjc08);济宁医学院青年基金项目(编号:TYQ2011K200J)

摘　　要：目的通过分子生物学方法构建小鼠Orexin 2β受体(mOX2βR)C末端突变表达载体。方法设计特异性引物,采用聚合酶链反应(PCR)技术,通过胶回收、酶切、连接、转化和测序等生物技术,构建mOX2βR突变体的表达载体。结果成功构建mOX2βR突变表达载体,将其分别命名为mu1、mu2和mu3。结论成功构建了mOX2βR C端不同的突变体,为进一步研究mOX2βR C末端氨基酸在信号通路中的作用奠定了基础。Objective To construct the mOX2βR mutation expression vector by using molecular biology meth- od. Methods We had designed specific primer in our research. These genes were amplified by Polymerase Chain Reaction（PCR）using the plasmid pcDNA3.1-mOX2βR as template. The PCR product was cut by restriction endonu- clease BamH I and Xbar I , connected by T4 DNA ligase and then be transformed into the E. coli competence DH5a. The construct of expression vector of mOX2βR mutants was identified by DNA sequencing. Results We successfully built mOX2βR mutant expression vector named mul, mu2 and mu3, respectively. Conclusion The mouse OX2βR mutant expression vectors were successfully constructed,and the expression vectors can be used to study the influence of mOX2βR carboxyl terminal mutations on the signal pathways,which will lay the foundation for the development of new drugs.

关 键 词：mOX2βR 突变体 G蛋白耦联受体 表达载体 

分 类 号：R37[医药卫生—病原生物学]