[Construction of a tRNAVal promoter plasmid expressing shRNAs in mammalian cells which mediates RNA interference].
OBJECTIVE
Constructing a plasmid containing tRNAVal promoter to express shRNA which mediates RNA interference.


METHODS
A tRNAVal gene was amplified from human genomic DNA by PCR and replaced the last several bases of 3' end by a linker. The tRNAVal promoter after artificial mutation followed a shRNA sequence to luciferase was cloned into pUC18, Puc-tRNAVal, lucRi Cotransfected with pMAMneoLuc into BHK-21 cell to detect the effect of luciferase expression.


RESULTS
pUC-tRNAVallucRi suppressed the luciferase expression from pMAMneoLuc by 97.9%-9.5%.


CONCLUSIONS
The results showed that the tRNAVal shRNA plasmid could efficiently suppress luciferase expression in BHK-21.
Lu
Xiao-chun
XC
Department of Geriatristic Cardiology, PLA General Hospital, Beijing 100853, China.
Yuan
Zhen-hua
ZH
Peng
Jian-qiang
JQ
Ma
Xin
X
Wu
Xiao-bing
XB
Li
Xiao-ying
XY
D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012343:RNA, Transfer / Q000235:genetics*
D012343:RNA, Transfer; D008156:Luciferases