中国畜牧兽医2012，Vol.39Issue(8)：44-47,4.
Arkadia表达载体转染肾小管上皮细胞条件的优化
Transfection Optimization of Arkadia Expression Vector in Renal Tubular Epithelial Cells
摘要
Abstract
To lay the foundation for investigating the role of Arkadia expression vector in mouse aristolochic acid nephropa-thy (AAN), we optimized the transfection conditions of ubiquitination enzyme Arkadia plasmid expression vector pGPU6/ GFP/Neo in mice renal tubular epithelial cells (RTECs). Mice RTECs were taken for transfection object, and liposome Lipo-fectamineTM 2000 for transfection medium, we detected the Arkadia Mrna expressions differences and the affects of cell activity under the different ratios conditions of plasmid and liposome by Real-time PCR and CCK-8 assay: at the same time, the differences of Arkadia Mrna expressions silence under the different transfection time conditions by Real-time PCR. The results showed that transfection efficiency of Arkadia-Pgpu6/GFP/Neo vector displayed dose and time dependents. When the ratio of plasmid DNA : LipofectamineTM 2000 was 0. 66 μg : 2 Μl, and the transfection time was 24 h, Arkadia Mrna expression inhibition was very obvious and the cytotoxicity was the lowest. Lastly, we determined the best transfection condition of Arkadia-Pgpu6/GFP/Neo vector.关键词
Arkadia/表达载体/转染/肾小管上皮细胞/优化Key words
Arkadia / expression vector / transfection/ renal tubular epithelial cells /optimization分类
生物科学引用本文复制引用
廖芳芳,李娇,王惠川,张中文,吴国娟..Arkadia表达载体转染肾小管上皮细胞条件的优化[J].中国畜牧兽医,2012,39(8):44-47,4.基金项目
北京市自然科学基金项目(6092004、51022014) （6092004、51022014）
国家自然科学基金(31172362、30972215) （31172362、30972215）
北京市属高校人才强教计划资助项目(PHR). （PHR）
