中国医科大学学报2011，Vol.40Issue(12)：1081-1084,4.DOI:21-1227/R.20111226.1615.028
NK4基因重组慢病毒载体的构建及在肝癌细胞中的表达
Construction of NK4 Recombinant Lentiviral Vector and Its Expression in HCCLM3 Cells
摘要
Abstract
Objective To study the transfection and expression level of human NK4 gene in HCCLM3 by constructing NK4 and EGFP fuse gene of recombinant lentiviral vector. Methods The NK4 gene was cloned to lentiviral expression vector with EGFP[pLenti6.3-IRES-EGFP]by recombining DNA technology .The 293T cells were cotransfected with lentiviral packaged systems and NK4 gene plasmid by lipo- fectin regeant to package the lentiviral particles, and the viral titer was determined.The 293T cells were infected with recombinant lentivirus by different multiple of infection(MOI). The best MOI was screened with reporter gene EGFP expression.HCCLM3 cells were infected by recombinant lentivirus at the best MOI.The transfection efficiency was assessed under fluorescent microscope and NK4 protein expression levels in transfected cells were detected by Western blot method. Results The recombinant lentiviral vector of NK4 gene and EGFP gene (pLen-ti6.3-NK4-IRES-EGFP) were successfully constructed.After being packaged, purified and concentrated, the virus reached a titer of 1.08X108 TU/mL.The best MOI was 7.There was significantly expression of NK4 protein in the transfected cells, but no expression in nontranfection cells. Conclusion The NK4 recombinant lentiviral vector had been successfully constructed and effectively transfected HCCLM3 cells/The NK4 protein was highly expressed in transfected HCCLM3 cells.关键词
NK4/慢病毒载体/人高侵袭转移肝癌细胞LM3/基因转染Key words
NK4/ lentiviral vector/ HCCLM3 cell/ gene transfection分类
医药卫生引用本文复制引用
李霏,李松林,尹元琴..NK4基因重组慢病毒载体的构建及在肝癌细胞中的表达[J].中国医科大学学报,2011,40(12):1081-1084,4.基金项目
辽宁省科技厅基金资助项目(2009225008-13) （2009225008-13）
