Construction of a tRNAVal promoter plasmid expressing shRNAs in mammalian cells which mediates RNA interference.
- Author:
Xiao-chun LU
1
;
Zhen-hua YUAN
;
Jian-qiang PENG
;
Xin MA
;
Xiao-bing WU
;
Xiao-ying LI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Line; Genetic Vectors; Humans; Luciferases; biosynthesis; genetics; Plasmids; genetics; Promoter Regions, Genetic; RNA Interference; RNA, Transfer; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2003;17(4):358-360
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEConstructing a plasmid containing tRNAVal promoter to express shRNA which mediates RNA interference.
METHODSA 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.
RESULTSpUC-tRNAVallucRi suppressed the luciferase expression from pMAMneoLuc by 97.9%-9.5%.
CONCLUSIONThe results showed that the tRNAVal shRNA plasmid could efficiently suppress luciferase expression in BHK-21.
