Arkadia表达载体转染肾小管上皮细胞条件的优化

Abstract

Abstract: To lay the foundation for investigating the role of Arkadia expression vector in mouse aristolochic acid nephropathy (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 Lipofectamine^TM 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∶Lipofectamine^TM 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.

Key words: Arkadia; expression vector; transfection; renal tubular epithelial cells; optimization

CLC Number:

Q813

LIAO Fang-fang, LI Jiao, WANG Hui-chuan, ZHANG Zhong-wen, WU Guo-juan. Transfection Optimization of Arkadia Expression Vector in Renal Tubular Epithelial Cells[J]. China Animal Husbandry & Veterinary Medicine.

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Transfection Optimization of Arkadia Expression Vector in Renal Tubular Epithelial Cells

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