基础医学与临床2011，Vol.31Issue(4)：440-444,5.
人NSPc1慢病毒过表达系统的建立与鉴定
Construction and identification of lentiviral over-expression system of human NSPcl gene
摘要
Abstract
Objective To construct and identify lentiviral over-expression system to study the function of NSPcl using over-expression technique. Methods The cDNA sequence of human NSPcl was chosen to design the primer,and restricted enzyme site was added to primer sequences. The DNA purified from PCR was digested with double restriction enzymes and then linked with linearized pLenti6-TO-EGFP-TRIP. The recombinants were identified by restriction digestion and DNA sequencing. The plasmids of positive clones were transfected into 293T cells, and Western blot was used to identify the over-expression efficiency of the lentiviral vector. Then 293T cells were used to package the lentiviral particles and infected by obtained lentiviral particles. The over-expression efficiency of NSPcl lentiviral system was examined by Western blot. Results The human NSPcl sequences was successfully inserted into pLenti6-TO-EGFP-TRIP vector, which was identified by double restriction digestion and DNA sequencing.Western blot validated that both the lentiviral vector and lentiviral particle can effectively over-express human NSPcl gene in 293T cells. Conclusion The construction of over-expression lentiviral system of human NSPcl gene may support the further study of function of NSPcl gene as a potential over-expression technique.关键词
过表达/NSPc1/慢病毒载体/多梳家族Key words
over-expression/ NSPcl/ lentivirus vector/ polycomb group分类
生物科学引用本文复制引用
李辉,龚燕华,胡光宇,阴彬..人NSPc1慢病毒过表达系统的建立与鉴定[J].基础医学与临床,2011,31(4):440-444,5.基金项目
国家自然科学基金(30971614,31070929) （30971614,31070929）
医学分子生物学国家重点实验室外部课题(2010) （2010）
