title: [Gene expression of methylene tetrahydrofolate reductase and its silencing effect in primary culture mouse embryo palatal mesenchymal cell].
abstract: OBJECTIVE
  To construct a small interfering RNA (siRNA) eukaryotic expression vector specific for methylene tetrahydrofolate reductase (MTHFR) gene and to observe its silencing effect on MTHFR gene.
  
  
  METHODS
  The expression vectors of PsiRNA-MTHFR were constructed by gene recombination and then were nucleofected into the primary cultured MEPM cell. At 48 h and 5 d after nucleofection, the expression of MTHFR in the levels of mRNA and protein was detected by real-time quantitative polymerase chain reaction (Real-Time PCR) and Western blot.
  
  
  RESULTS
  The eukaryotic expression vector of PsiRNA-MTHFR, which significantly down-regulated mRNA and protein of MTHFR at 48 h and 5 d after nucleofection, were successfully constructed.
  
  
  CONCLUSIONS
  Eukaryotic expression vector of siRNA specific for MTHFR is successfully contructed, which lays the basis for its application in the mechanism research of MTHFR gene regulating embryo palate shelves fusion.
authors.affiliation: Dept. of Oral and Maxillofacial Surgery, The Medical School Hospital of Qingdao University, Qingdao 266003, China.
authors.forename: Wen-Lin
authors.initials: WL
authors.lastname: Xiao
authors.forename: Bing
authors.initials: B
authors.lastname: Shi
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Qian
authors.initials: Q
authors.lastname: Zheng
authors.forename: Lei
authors.initials: L
authors.lastname: Huang
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D042965:Methylenetetrahydrofolate Reductase (NADPH2); D051379:Mice; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D042965:Methylenetetrahydrofolate Reductase (NADPH2)