Biotechnology Bulletin ›› 2013, Vol. 0 ›› Issue (10): 109-112.

• Report • Previous Articles     Next Articles

Construction and Analysis of Luciferase Reporter Vectors of Mouse Dnmt1 Gene Promoter

Zhang Yuan1,Yang Xunxun2, Wu Fengrui2,，3, Liu Yong2,，3, Ding Biao2,，3, Wang Caihong2, Huang Jichang2, Li Wenyong2,，3,   

  1. （1. College of Life Science，Anhui Agricultural University，Hefei 230036 ；2. Key Laboratory of Embryo Development and Reproductive Regulation，Anhui Province，Fuyang 236037 ；3. College of Life Science，Fuyang Normal University，Fuyang 236037）
  • Received:2013-04-11 Revised:2013-10-15 Online:2013-10-14 Published:2013-10-15

PDF (PC)

1366

Abstract:

To study the different 5'-flanking regions activity of mouse Dnmt1 gene, 4 truncated Dnmt1 promoters were amplified by polymerase chain reaction（PCR）from mouse genomic DNA and subcloned into pMD19-T vector, and then cloned into the pGL3-Basic luciferase reporter gene vector to construct Dnmt1 promoter-pGL3-Basic recombinants. After the recombinants were transfected into NIH/3T3 cells with Lipofectamine, relative luciferase activity were obtained by using the Dual-Luciferase Reporter Assay System and Glomax. Results showed that the recombinants of 5'-flanking sequential deletion of mouse Dnmt1 promoter-pGL3-Basic reporter gene were successfully constructed. The relative luciferase activity of Dnmt1-1-pGL3-Basic was three times higher than the other three truncated constructs. Together, this study was preliminary confirmed that the mouse Dnmt1 promoter region（-1 866-+57 bp）had strong transcriptional activity.

Key words: Mouse, Dnmt1 promoter, NIH/3T3 cell line, Activity analysis