ClC-3荧光真核表达载体的构建及表达  

Construction and expression of ClC-3 gene fluorescent eukaryotic cell expression vector

作　　者：杨清松[1,2] 张守盼[1] 林卓华[1] 毛建文[3] 徐彬[1] 

机构地区：[1]广东药学院生命科学与生物制药学院,广州510006 [2]中国科学院大学,北京100049 [3]广东药学院基础学院,广州510006

出　　处：《中国当代医药》2013年第29期4-5,8,共3页China Modern Medicine

基　　金：国家自然科学基金(81101666);广东药学院大学生创新实验项目(75)

摘　　要：目的构建表达ClC-3的荧光真核表达载体,为研究其在肿瘤细胞中的作用提供分子工具。方法将ClC-3的编码序列亚克隆到pEGFP-N1质粒,构建荧光真核表达载体pEGFP-N1/ClC-3,利用脂质体介导将ClC-3基因导入人乳腺癌细胞MCF-7内,通过绿色荧光观察检测基因的表达。结果成功构建荧光真核表达载体pEGFP-N1/ClC-3,该载体转染MCF-7细胞后,可观察到绿色荧光。结论表达ClC-3的荧光真核表达载体构建成功,并在乳腺癌细胞中得到正确表达,为进一步研究ClC-3在乳腺癌中的作用奠定了基础。Objective To construct fluorescent eukaryotic cell expression vector carrying the gene of C1C-3 and pro- vide the molecular tools for further study of its role in tumor. Methods The C1C-3 coding sequence was subcloned in- to pEGFP-N1 plasmid to construct fluorescent eukaryotic expression vector pEGFP-NI/CIC-3.pEGFP-N1/C1C-3 was transfected into MCF-7 cells by liposome mediating and gene expression was detected by green fluorescent observa- tion. Results pEGFP-N1/C1C-3 was successfully constructed,specific green fluorescence was detected by fluorescence microscope after the recombinant vector transfected. Conclusion The fluorescent eukaryotie expression vector coding for C1C-3 is successfully constructed and correctly expressed in breast carcinoma cells.This investigation provides the basis for further study of its action in breast cancer.

关 键 词：CLC-3 MCF-7细胞 荧光真核表达载体 转染 

分 类 号：R737.9[医药卫生—肿瘤] R453[医药卫生—临床医学]