Cloning and Expression of Influenza H1N1 NS1 Protein in <italic>Escherichia Coli</italic> BL21

Abstract

Background:Influenza virus is globally pathogenic and it is usually associated with zoonotic respiratory diseases. This virus has caused a number of pandemics with a high mortality rate. The non-structural (NS1) protein of influenza A viruses is a non-essential virulence factor that has multiple accessory functions during viral infection. This protein is highly conservative. It has been shown that this protein has a major role against the immunity responses of the host cells. Objectives:The aim of this study is to produce the recombinant Influenza NS1 protein by the use of bacterial production system in order to evaluate the immunological and structural features of the protein in the following researches. Materials and Methods:The NS1 gene construct has been artificially synthesized; subsequently it has been sub-cloned in to the pQE30 expression vector. Then the expression vector has been transformed in to the BL21 cells and induced by IPTG, afterward the expression has been evaluated by SDS-PAGE and Western Blotting Techniques. Results:The NS1 gene has successfully cloned and transformed in to expression cells, as a result a 23 kDa band has been observed both on 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 bacterial protein translation system. Science this protein is a conservative protein among Influenza A viruses could be used as a potent vaccine for prevention of various types of pandemics caused by Influenza A.