摘要
目的:克隆大鼠促甲状腺激素释放激素受体1(TRH-R1)基因,构建其真核表达载体,并检测该基因在非洲绿猴肾细胞系COS-7中的表达。方法:应用RT-PCR方法,以大鼠脑源RNA为模板,扩增获得TRH-R1基因,定向克隆到pDsRed2-N1中,以LipofectAMINE 2000试剂转染pDsRed2-N1-TRH-R1表达载体至COS-7细胞系中进行瞬时表达。结果:测序结果表明,从大鼠脑源总RNA中克隆到正确的TRH-R1基因全长编码序列;显微照相观察到所构建的TRH-R1表达载体质粒在COS-7细胞系中获得有效表达。结论:大鼠TRH-R1基因的克隆、真核表达载体的构建及在COS-7细胞系中表达获得成功,为进一步研究其功能奠定了基础。
Objective：To clone rat thyrotropin releasing hormone receptor 1（TRH-R1） gene and express it in Africa green monkey kidney COS-7 cell lines.Methods：TRH-R1 gene was amplified from RNA of rat brain tissue by RT-PCR,and then it was inserted into eukaryotic expression vector pDsRed2-N1.The recombinant expression vector was transiently transfected into COS-7 cell lines by LipofectAMINE 2000 reagent.Results：The sequence of the cloned TRH-R1 was confirmed to be correct by DNA sequencing.The expression of TRH-R1 in COS-7 cell lines was detected by fluorescence microscope,and the pictures showed that the TRH-R1 protein was expressed efficiently.Conclusion：The cloning of rat TRH-R1 gene and the construction of its eukaryotic expression vector were finished,which lays the foundation for further investigating the role of TRH-R1
出处
《生物技术通讯》
CAS
2011年第1期19-23,共5页
Letters in Biotechnology
