中国医科大学学报2012，Vol.41Issue(2)：112-114,119,4.
转录因子Kaiso真核表达载体的构建及鉴定
Construction and Identification of Kaiso Eukaryotic Expression Vector
摘要
Abstract
Objective To study the biological function of transcription factor Kaiso in the lung cancer cells and construct the eukaiyotic expression vector of human Kaiso gene. Methods Kaiso Cdna was amplified by PCR and inserted into eukaryotic expression vector Pegfp. The construction of the recombinant Pegfp-Kaiso was tested by restriction enzyme analysis and gene sequencing . Then ihe Pegfp-Kaiso was transfected into lung cancer cell lines,A549 cells. Last,the localization and expression of Kaiso in bansfected A549 cells were verified by fluorescent microscope and Western blot,respectively. Results The size of inserted fragments in the recombinant vectors Pegfp- Kaiso is 2 022 bp, corresponding to that of Kaiso. After transfecting Pegfp-Kaiso into A549 cells by using IipofectamineTM 2000, The CFP signal was detected in the nucleus and cytoplasm,while the expression level of Kaiso in transfected A549 cells was significantly elevated (P < 0.05). Conclusion The eukaryotic expression vector Pegfp-Kaiso is successfully constructed and it provides a powerful tool for studying the biological function of transcription factor Kaiso.关键词
Kaiso/基因克隆/真核表达载体Key words
Kaiso/ gene cloning/ eukaiyotic expression vector分类
医药卫生引用本文复制引用
曹红一,唐娜,林旭勇,张迪,苗原,王恩华,戴顺东..转录因子Kaiso真核表达载体的构建及鉴定[J].中国医科大学学报,2012,41(2):112-114,119,4.基金项目
国家自然科学基金资助项目(81071717,81000942) （81071717,81000942）
