中德临床肿瘤学杂志（英文版）2006，Vol.5Issue(2)：141-143,3.
B7-1/GFP基因真核表达载体的构建及其在骨肉瘤细胞中的表达
Construction of Eukaryotic Expression Vector Containing B7-1/GFP Gene and Its Expression in Osteosarcoma Cell Line
摘要
Abstract
Objective: To construct eukaryotic expression vector containing B7-1/GFP geneand study its expression in osteosarcoma cell line LM8. Methods: By using gene cloning technique, eukaryotic expression vector pEGFP C1 was used to construct the murine B7-1 recombinant plasmid (pEGFP C1/B7). Recombinant plasmid was transfected into LM8 cells with liposome and was confirmed by restriction endonuclease digestion and DNA sequencing. The expression of the fusion protein was detected using fluorescence microscope and Western blot analysis. Results: The recombinant eukaryotic expression plasmid pEGFP-C1/B7 was successfully constructed, which was confirmed by DNA sequencing, RT-PCR and restriction enzymes analysis. The green fluorescent protein could be detected in the transfected LM8 with fluorescence microscope. The expected B7-1 and green fluorescent protein (GFP) fusion protein was detected by RT-PCR and Western blot. Conclusion: The eukaryotic expression vector containing B7-1/GFP gene was constructed successfully, and it could be expressed in LM8 after transfection.关键词
B7基因/绿色荧光蛋白/基因重组/骨肉瘤Key words
B7-1 gene/green fluorescent protein/gene recombination/osteosarcoma分类
医药卫生引用本文复制引用
宁旭,刘勇,杨述华,傅德皓..B7-1/GFP基因真核表达载体的构建及其在骨肉瘤细胞中的表达[J].中德临床肿瘤学杂志（英文版）,2006,5(2):141-143,3.基金项目
This study was supported by a grant from the youth dawn program of Wuhan (Grant No. 20025001028). （Grant No. 20025001028）
