Construction of NF-kappaB siRNA expression vector.
- Author:
Yunusi KUREXI
1
;
Guo-hong HUANG
;
Mei LONG
;
Jie HE
;
Maimaitiaili NASIER
;
Yan-ping XIA
;
Wufuer HAMULATI
Author Information
1. Department of Biochemistry and Molecular Biology, Xinjiang Medical University, Urumqi 830054, China.
- Publication Type:Journal Article
- MeSH:
Animals;
COS Cells;
Chlorocebus aethiops;
Genetic Vectors;
NF-kappa B;
genetics;
RNA Interference;
RNA, Small Interfering;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2007;32(5):765-770
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct a NF-kappaB siRNA expression vector and to detect the specific silencing effect of the siRNA on the expression of NF-kappaB protein.
METHODS:pcDNA3.1/CT-GFP-TOPO recombinant eukaryotic expression vector and pSilencer 1.0-U6-siRNA-NF-kappaB recombinant vector were constructed respectively. These 2 recombination plasmids were co-transfected into COS-7 cells, and the NF-kappaB silence induced by RNAi was detected by Western blot and the inverted fluorescence microscope.
RESULTS:The levels of NF-kappaB protein in COS-7 cells could be silenced effectively and specifically by pSilencer 1.0-U6-siRNA- NF-kappa recombinant vector. The expression of NF-kappaB protein was reduced gradually with the increase of pSilencer 1.0-U6-siRNA- NF-kappaB recombinant vector,which could be detected by Western blot under the inverted fluorescence microscope.
CONCLUSION:NF-kappaB siRNA expression vector is constructed successfully.
