siRNA对猪肝细胞THRSP基因mRNA表达的干扰效率.PDFVIP

安徽农业大学学报, 2014, 41(3): 363-366 Journal of Anhui Agricultural University [DOI] 10.13610/ki.1672-352x014 网络出版时间：2014-4-23 15:03:00 [URL]/kcms/doi/10.13610/ki.1672-352x014.html siRNA 对猪肝细胞 THRSP 基因 mRNA 表达的干扰效率 1 1 1, 2* 1 1 1 1 1 潘中婷 ，王学故 ，陈宏权 ，焦明慧 ，谢亚男 ，周 梅 ，陈公伟 ，马帮军 (1. 安徽农业大学动物科技学院，合肥 230036 ；2. 安徽地方畜禽遗传资源保护与生物育种省级实验室，合肥 230036) 摘 要：应用改良的剪切消化法（1%Ⅳ型胶原酶）消化分离仔猪肝脏细胞，建立稳定的仔猪肝脏细胞的分离 与原代培养体系；通过siRNA 途径及RT-PCR 技术检测肝细胞THRSP 基因mRNA 表达水平。结果表明，平均每g 6 肝脏可获得6.2 ×10 个细胞，接种即时存活率达94.84% ，培养存活率达50%左右，细胞转染后细胞存活率达80% 以上；设计合成的siRNA 的干扰效率为75.3% 关键词：猪；肝脏细胞原代培养；siRNA；THRSP 基因；mRNA 中图分类号：S828.2 文献标识码：A 文章编号：1672−352X (2014)03−0363−04 Interfered efficiency of siRNA on mRNA expression of THRSP gene in pig liver cells 1 1 1, 2 1 PAN Zhongting , WANG Xuegu , CHEN Hongquan , JIAO Minghui , 1 1 1 1 XIE Yanan , ZHOU Mei , CHEN Gongwei , MA Bangjun (1. School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036; 2. Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Bio-breeding, Hefei 230036) Abstract: In this study, digesting and separating piglets liver cells obtained from modified shear digestion (1% Ⅳcollagenase )were cultured and dynamically observed with inverted microscopy, so as to establish a stable system for primary culture of piglets liver cells. RNAi and RT-PCR technique were employed to the detection of the cell mRNA expression level, and then the changes of THRSP mRNA expression at the cellular level were ex- 6 plored. The results showed that 6.2×10