The cloning of mouse nicotinamide mononucleotide adenylyl-transferase gene and the detecting of its expression / 中华老年医学杂志
Chinese Journal of Geriatrics
;
(12): 866-868, 2011.
Article
in Chinese
| WPRIM
| ID: wpr-422599
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Geriatrics
Year:
2011
Type:
Article
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