安徽医科大学学报Issue(2)：145-148,4.
鼠源pEGFP-C2-NLRC5重组质粒的构建及其表达
Construction of recombinant pEGFP-C2-NLRC5 and its expression
摘要
Abstract
Objective To construct the GFP-tagged eukaryotie expression vector of NLRC5 and observe its expres-sion in mouse macrophage RAW264 . 7 . Methods The cDNA of NLRC5 was obtained from mouse macrophage RAW264. 7 cells, amplified by PCR and cut with double enzyme EcoR I and BamHI, then inserted into the eu-karyotic expression vetor pEGFP-C2. The recombinant vector was verified by PCR, restriction enzymes cut and se-quencing identified. Then transfected into mouse macrophage RAW264. 7 cells and the expression of pEGFP-C2-NLRC5 was monitored by fluorescence, PCR and Western blot. Results To deal with recombinant pEGFPC2-NL-RC5 with double digestion, then fragments of NLRC5 could be seen, also GFP could be detected in the transfected RAW264. 7 cells. NLRC5 gene expression could be detected by PCR,and its protein expression was detected by Western blot. Conclusion Eukaryotic expression vector of NLRC5 is successfully constructed, and the fusion ex-pression of NLRC5 protein and GFP can be detected in RAW264 . 7 .关键词
NLRC5/pEGFP-C2/RAW264.7/基因表达/蛋白表达Key words
NLRC5/pEGFP-C2/RAW264.7/gene expression/protein expression分类
医药卫生引用本文复制引用
李琳,彭云云,黄成,徐涛,李俊..鼠源pEGFP-C2-NLRC5重组质粒的构建及其表达[J].安徽医科大学学报,2014,(2):145-148,4.基金项目
国家自然科学基金(编号:81273526、81072686) （编号:81273526、81072686）
