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.
Animals ; COS Cells ; Chlorocebus aethiops ; Genetic Vectors ; NF-kappa B ; genetics ; RNA Interference ; RNA, Small Interfering ; Transfection