[Construction of an expression vector for high expression of siRNA].
OBJECTIVE
To construct the vector for efficient expression of siRNA using pre-mir30 backbone.


METHODS
By chemical synthesis method, pre-mir30 backbone introduced an appropriate restriction enzyme site for foreign shRNA inserting was cloned into an expressing vector containing U6 promoter. The silencing efficiency of a new siRNA expressing vector was detected by transfection and Western blot.


RESULTS
The new vector containing pre-mir30 backbone expressing siRNA against GFP could markedly inhibit the expression of GFP compared with the vector expressing control siRNA.


CONCLUSIONS
siRNA expressing vector constructed by pre-mir30 backbone could highly express foreign siRNA.
College of Life Sciences, Shaanxi Normal University, Xi'an 710100, China. lihuijin@stu.snnu.edu.cn
Hui-Jin
HJ
Li
Xiao-Jing
XJ
Zheng
Wei-Feng
WF
Zhang
Ye
Y
Bian
Dong-Yang
DY
Wang
Jun-Li
JL
Zhao
Hai-Bin
HB
Xia
D000818:Animals; D002417:Cattle; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000096:biosynthesis* / Q000235:genetics*
D035683:MicroRNAs; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D004262:DNA Restriction Enzymes