Construction of eukaryotic expression vectors of nephroblastoma overexpression gene and expression in rat dermal multipotent stem cells

VernacularTitle:NOV基因真核表达载体的构建及在大鼠真皮多能干细胞中的表达
Author: Yan-Ping LIU ¹ ; Lu-Si LI ; Tao WANG ; Wen-Yue XU ; Ji-Fu QU
Author Information

1. 第三军医大学附属西南医院
Keywords: Nephroblastoma overepression gene; RT-PCR; Gene cloning; DMSCs
From: Chinese Journal of Neuromedicine 2008;7(4):353-356
CountryChina
Language:Chinese
Abstract: Objective To obtain eukaryotic expression vectors containing coding region of nephroblastoma over-expression gene (NOV) and detect its expression in dermal multipotent stem cells(DMSCs). Methods A 1 178 bp cDNA fragment was amplified from the total RNA of normal rat brain tissue by RT-PCR and cloned into eukaryotic expression vector pEGFP-N1. The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes Hind Ⅲ and BamH Ⅰ.The recombinant plasmid was transfected into DMSCs with liposome. The expression of NOV gene was detected by RT-PCR. Results Eukaryotic expression vectors containing 1 178-bp coding region of NOV gene were successfully constructed. DMSCs transfected with the recombinant plasmid expressed NOV gene. Conclusions That eukaryotic expression vector containing coding region of NOV gene is constructed and expressed in DMSCs can provide a strong molecular tool for the studies on the NOV gene and DMSCs.