基础医学与临床Issue(9)：992-997,6.
EGFP-线粒体蛋白导入载体的构建及功能验证
Construction of the mitochondrial protein imported vector with EGFP and confirmation
摘要
Abstract
Objective To construct a eukaryotic expression vector which can transport foreign protein into mitochondria. Methods The mitochondrial transit peptide's sequences were fused with EGFP through multiplex PCR amplification and inserted into the eukaryotic expression vector pcDNA3. 1, constructing the eukaryotic expression vector pcDNA5. 1-EGFP. Then the P53 gene was inserted into pcDNA5. 1-EGFP and pcDNA3. 1, constructing pcDNA5. 1-P53 and pcDNA3. 1-P53 vector. After verification by restriction enzyme digestion and sequencing, the plasmids were transfected into 293T cells for profiling the distribution of EFGP and cytochrome C in the cell, and the positioning of P53 protein in the cell. Results The distribution of EGFP was consistent with cytochrome C, and the P53 protein expressed by pcDNA5. 1-P53 was concentrated in mitochondria in the cytoplasm, while the vector pcDNA3. 1-P53 was mainly distributed in the nucleus. Conclusions The eukaryotic expression vector was successfully constructed, which may transport foreign protein into the mitochondria.关键词
表达载体/绿色荧光蛋白/线粒体导肽Key words
expression vector/ green fluorescent protein/ mitochondrial transit peptide分类
医药卫生引用本文复制引用
乔录新,张玉林,丁渭,陈德喜..EGFP-线粒体蛋白导入载体的构建及功能验证[J].基础医学与临床,2012,(9):992-997,6.基金项目
国家"十一五"传染病重大专项(2008ZX10001-004,2008ZX10001-007) （2008ZX10001-004,2008ZX10001-007）
国家自然科学基金(30910103915,30870853) （30910103915,30870853）
