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[Construction of recombinant NF-κB reporter adenovirus and activity analysis]
Jin-hai Guo1, Feng Wang, Ping-zhang Wang
1Peking University School of Basic Medical Sciences, Beijing, China.
Objective:
To study the feasibility of adenovirus-based nuclear factor-κB (NF-κB) reporter as a model to screen the upstream signal regulators of NF-κB.
Methods:
A type 5 (E1/E3 deficient) adenovirus vector pAdxsi was used to construct the NF-κB reporter adenovirus. Multiple adherent and suspending cell lines were infected by the NF-κB reporter adenovirus, and the luciferase activity of the NF-κB reporter gene was measured.
Results:
An NF-κB reporter adenovirus (Ad-NF-κB-luc) was successfully constructed. The virus was capable of infecting HepG2, MGC803, THP-1 and U937 cell lines and showed high activities of NF-κB-luc reporter gene when stimulated by tumor necrosis factor-α (TNF-α) or lipopolysaccharide (LPS).
Conclusion:
The Ad-NF-κB-luc reporter gene transfer system can effectively infect those cells hard-transfected by conventional transfection reagents. It also produces a high activity of NF-κB-luc reporter gene with stability and reliability. Our study expands the application of NF-κB reporter gene.
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