期刊论文

Construction and Subcellular Localization of SDF-1α -incorporating GFP Fusion Protein Vector

中文

中国比较医学杂志

1671-7856

2008

18

3

1-4+83

10.3969/j.issn.1671-7856.2008.03.001

湖南省自然科学基金（06JJ50051） 湖南省教育厅科学研究项目（06C693）.

本校为第一机构

目的 构建SDF-α基因与绿色荧光蛋白的融合蛋白表达载体,进而观察SDF-1α基因编码蛋白在细胞内的定位情况.方法 用EcoRI内切酶从pMD-T18-SDF-1α重组载体中酶切分离SDF-α基因的完整ORF,构建pEGFP-C1-SDF-lα的融合表达载体,脂质体转染COS-7细胞,并在荧光显微镜下观察表达的融合蛋白.结果 SDF-lα基因在COS-7细胞中高效表达,激光共聚焦的结果显示,SDF-1α基因定位在细胞质内.结论 成功构建了pEGFP-C1-SDF-1α的融合表达载体,SDF-lα基因主要在细胞质中表达.

Objective To construct the SDF-1α-incorporating GFP fusion protein vector, and to observe the subcellular fraction of its encoding protein. Methods The complete coding sequence of SDF-1α gene was obtain from pMD-T18-SDF-1α recombinant plasmid by EcoRI restriction enzyme digestion, and then clued into pEGFP-C1 on special direction. After these recombinant plasmids were transfected into COS-7 cell line by liposome, the expression of SDF-1α gene and its subcellular fraction were observed by fluorescent microscope and laser confocal microscope....