小鼠Myf5真核表达载体的构建及其在细胞中的定位  

Construction of the Eukaryotic Expression Vector Carrying Encoding Gene of Mouse Myf 5 and Its Location in 10T1/2 Cells

作　　者：刘向东[1] 郭芬[1,2] 黄晓峰[1] 刘兆宇[1] 张欣[1] 李月琴[1] 周天鸿[1] 

机构地区：[1]暨南大学生命科学技术学院,广东广州510632 [2]广东药学院生命科学与生物制药学院,广东广州510006

出　　处：《生物技术》2010年第3期11-13,共3页Biotechnology

基　　金：高等学校博士学科点专项科研基金项目(No20060559006)资助

摘　　要：目的:获得小鼠生肌调节因子Myf5基因并构建pEYFP-C1真核表达载体,观察Myf5在小鼠C3H10T1/2细胞中的定位。方法:利用PCR获得Myf5基因克隆到pEYFP-C1载体中,利用脂质体将构建的表达载体转染C3H10T1/2细胞,荧光观察融合蛋白的表达。结果:从小鼠cDNA文库中得到760bp的myf5的CDS序列后,重组到pEYFP-C1载体中并转染C3H10T1/2细胞,荧光显示Myf5蛋白定位在细胞核中。结论:Myf5载体成功构建并在小鼠C3H10T1/2细胞表达,证明了Myf5蛋白定位于细胞核,为进一步研究Myf5与其他蛋白的相互作用奠定了基础。Objective：To construct the recombinant eukaryotic expression vector pEYFP-C1 /Myf5 carrying encoding gene of mouse Myf5 and test its location in 10T1 /2 cells.Method：The full length of Myf5 cDNA sequence was amplified from mouse cDNA library by PCR.Then Myf5 CDS fragment was ligated into plasmid pEYFP-C1.C3H10T1 /2 cells were transfected with pEYFP-C1 /Myf5 vector.The expression of Myf5 was observed by fluorescence microscope.Result：Myf5 CDS which was about 760bp and obtained by PCR was recombinant into pEYFP-C1 voctor.C3H10T1 /2 cells were transfected with pEYFP-C1 /Myf5 vector and Myf5 protein was found locating in nucleus.Conclusion：The eukaryotic expression plasmid pEYFP-C1 /Myf5 has been successfully constructed and expressed in the C3H10T1 /2 cells.Myf5 protein has been proved locating in nucleus.

关 键 词：MYF5 C3H10T1/2 细胞定位 

分 类 号：Q786[生物学—分子生物学]