广西医学2012，Vol.34Issue(6)：677-680,4.
IGF-Ⅱ基因RNAi慢病毒载体构建与稳定转染细胞株建立
Construction of IGF-Ⅱ RNA Interference Lentiviral Vector and Establishment of Its Stably Transfected Cell Line
摘要
Abstract
Objective To construct the IGF- Ⅱ RNA interference lentiviral vector and establish the stably transfected human adrenocortical tumor cell line SW-13. Methods According to the IGF-Ⅱ gene sequence,three shRNA targeting IGF- II were designed, and the corresponding pLLU2G-shIGF recombinant plasmids were constructed. After the most effective shRNA was achieved, the shuttle plasmid pLLU2G-shIGF with lentiviral packaging plasmid was mixed,and 293FT cells were transfected. Virus solution was collected to infect SW-13 cells,and the stably transfected cell line was established. Results The third shRNA sequence targeting IGF- Ⅱ showed the best gene silencing effect as 80% , and the stable transfected cell line was constructed successfully based on this sequence, and the inhibition ratio was 95% for IGF- ⅡmRNA. Conclusion It is successful to obtain IGF-Ⅱ-shRNA lentiviral vector that can effectively down-regulate the expression of IGF- Ⅱ in human adrenocortical tumor cell line SW-13 and establish the stably transfected cell line.关键词
IGF-Ⅱ/RNAi/SW-13/shRNA/慢病毒载体/转染细胞Key words
IGF- Ⅱ/ RNAi/ SW-13/ shRNA/ Lentiviral vector/ Transfected cell分类
医药卫生引用本文复制引用
杨翠英,罗佐杰..IGF-Ⅱ基因RNAi慢病毒载体构建与稳定转染细胞株建立[J].广西医学,2012,34(6):677-680,4.基金项目
中华医学会内分泌学分会边远地区基金项目(CSE2009009) （CSE2009009）
