[Gene expression of methylene tetrahydrofolate reductase and its silencing effect in primary culture mouse embryo palatal mesenchymal cell].
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.
Xiao
Wen-Lin
WL
Dept. of Oral and Maxillofacial Surgery, The Medical School Hospital of Qingdao University, Qingdao 266003, China.
Shi
Bing
B
Wang
Yan
Y
Zheng
Qian
Q
Huang
Lei
L
D000818:Animals; D002478:Cells, Cultured; D042965:Methylenetetrahydrofolate Reductase (NADPH2); D051379:Mice; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014162:Transfection
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D042965:Methylenetetrahydrofolate Reductase (NADPH2)