山东医药2011，Vol.51Issue(33)：16-18,3.
细胞外基质金属蛋白酶诱导因子RNAi慢病毒载体的构建
Construction of lentiviral vector of EMMPRIN RNAi
摘要
Abstract
Objective To construct lentivirus vector for RNA interference( RNAi) targeting extracellular matrix metal-loproteinase inducer(EMMPRIN). Methods RNAi targeting sequence was designed. The complementary Oligo DNA targeting EMMPRIN were synthesized, and then forming double strand DNA after annealing, connecting pGCSIL-GFP vector after enzyme digestion by Age I and EcoR I , forming lentivirus vector pGC-LV. The positive clones containing Emmprin shRNA were identified by PCR amplification and sequencing. 293 T cells were cotransfected with lentiviral vector pGC-LV, pHelper 1.0 and pHelper 2.0 using Lipfectamine 2000. The recombinant lentivirus were collected and detected by dilution method one hole by another. Results A recombinant lentiviral vector which expressed short hairpin RNA targeting Emmprin was successfully constructed and confirmed by DNA sequencing. The recombinant lentivirus was harvested from 293T cells and the viral liter was 1 × 109 TU/ml. Conclusion RNAi lentivirus vector of EMMPRIN was constructed successfully, and the vector provides a basis for further studying of the roles of Emmprin in acute leukemia.关键词
细胞外基质金属蛋白酶诱导因子/RNA干扰/慢病毒载体Key words
extracellular matrix metalloproteinase inducer/ RNA interference/ lentiviral vector分类
医药卫生引用本文复制引用
涂燕,李振江,汤爱萍,李慧慧,贺文凤,李洁,张冉..细胞外基质金属蛋白酶诱导因子RNAi慢病毒载体的构建[J].山东医药,2011,51(33):16-18,3.基金项目
国家自然科学基金(30800490) （30800490）
江西省青年科学家(井冈之星)培养对象项目(2009DQ02000). （井冈之星）
