The cloning of mouse nicotinamide mononucleotide adenylyl-transferase gene and the detecting of its expression
10.3760/cma.j.issn.0254-9026.2011.10.021
- VernacularTitle:小鼠烟酰胺单核苷酸腺苷酰转移酶1基因的克隆与表达检测
- Author:
Hong ZHAO
;
Zichao YANG
;
Jingyu ZHANG
- Publication Type:Journal Article
- Keywords:
Sulfate adnylyl transferase;
Gene expression
- From:
Chinese Journal of Geriatrics
2011;30(10):866-868
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct eukaryotic expressing vector of the mouse NMNAT1 (nicotinamide mononucleotide adenylyl-transferase) gene and examine its ability to express the NMNAT1 gene in Hela cells.Methods The full-length NMNAT1 cDNA sequence was amplified by PCR and cloned into the plasmid of T-vector and then to pcDNA3.1 construct.The recombinant plasmid pcDNA3.1-NMNAT1 was identified by DNA sequencing and then transfected with Lipofectamine2000 into Hela cells.The expression of NMNAT1 was detected by real time quantitative PCR (qPCR) and Western blot after 48 h transfection.Results The recombinant eukaryotic vector carrying NMNAT1 gene was constructed successfully in a match with database and this vector could up-regulate the expression of the NMNAT1 gene both in mRNA and protein levels in Hela cells.Conclusions The eukaryotic vector carrying NMNAT1 gene (pcDNA3.1-NMNAT1) enhances the expression of NMNAT1 gene.
