Cloning and expression of influenza H1N1 NS1 protein in Escherichia Coli BL21
MetadataShow full item record
Background: The influenza virus is globally pathogenic and it is usually associated with zoonotic respiratory diseases. This virus has caused a number of pandemics with high mortality rates. The non-structural protein-1 (NS1) of influenza A viruses is a non-essential virulence factor that has multiple accessory functions during a viral infection. This protein is highly conservative. It has been shown that this protein plays a major role against the immunity responses of host cells. Objectives: The aim of this study was to produce the recombinant influenza NS1 protein by the use of a bacterial production system, in order to evaluate the immunological and structural features of this protein in future researches. Materials and Methods: The NS1 gene construct was artificially synthesized; subsequently it was sub-cloned into the pQE30 expression vector. The expression vector was then transformed into the BL21 cells and induced by IPTG. Finally, the expression was evaluated by SDS-PAGE and Western blotting techniques. Results: The NS1 gene was successfully cloned and transformed into expression cells. As a result, a 23 kDa band was observed both on the SDS-PAGE and nitrocellulose paper after Western blotting. Conclusions: Based on the results of this study, it could be concluded that the NS1 gene of influenza A H1N1 virus (A/Shiraz/14/2010 strain) could be cloned and the rNS1 protein (recombinant NS1 protein) could be expressed using a bacterial protein translation system. Since this protein is a conservative protein among influenza A viruses, it could be used as a potent vaccine for the prevention of various types of pandemics caused by influenza A. © 2014, National Institute of Genetic Engineering and Biotechnology; Published by Kowsar Corp.