中国医科大学学报2011，Vol.40Issue(8)：684-687,4.DOI:21-1227/R.20110726.1659.024
小鼠3D3/LYRIC基因重组质粒构建及蛋白表达和定位
Construction of Fusion Plasmid of Mouse 3D3/LYRIC Gene and Identification of Its Recombinant Protein Expression and Localization
摘要
Abstract
Objective To construct eukaryotic expression plasmid of mouse 3D3/LYRIC gene and identify its recombinant protein expression and localization. Methods Total mRNA isolated for a mouse mammary epithelial C127 cells was reversely transcribed into cDNA. The m3D3/LYRIC coding sequence was amplified by polymerase chain reaction (PCR) method and was subcloned into pEGFP-C1 expression vectors. After the target region was sequenced,the plasmids were transfected into ACC-2 cells,which were established from adenoid cystic carcinoma cells of the salivary gland (ACC-2 cells) . Expression of the recombinant plasmids in ACC-2 cells was detected by Western blot. The localization of pEGFP-m3D3/LYRIC in ACC-2 cells was observed by laser scanning confocal microscopy. Results m3D3/LYRIC has been constructed into eukaryotic expressing vector pEGFP-Cl successfully. The length of the fragment was 1740 bp,identified by restriction enzymes digestion. The expression of GFP-m3D3/LYRIC fusion protein was detected by Western blot,with a molecular weight of 91 kDa. The pEGFP-m3D3/LYRIC protein was mainly localized in the cytoplasma without any expression in the nucleus. Conclusion The recombinant plasmids were successfully cloned into eukaryotic expressing vector m3D3/LYRIC. The pEGFP-m3D3/LYRIC fusion protein was mainly expressed in the cytoplasma.关键词
3D3/LYRIC/腺样囊性癌/AEG-1Key words
3D3/LYRIC/ adenoid cystic carcinoma/ AEG-1分类
医药卫生引用本文复制引用
代炜,李彦姝,李妍,孙长伏..小鼠3D3/LYRIC基因重组质粒构建及蛋白表达和定位[J].中国医科大学学报,2011,40(8):684-687,4.基金项目
国家自然科学基金资助项目(30672331,30800415) （30672331,30800415）
