Construction of NF-kappaB siRNA expression vector / 中南大学学报(医学版)

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.