Cloning of influenza B NS1 gene in Escherichia Coli

Abstract

The non-structural NS1 protein of influenza B virus is a multifunctional virulence protein which is involved in the transport of viral RNA. Inside the host cell, NS1B antagonizes and inhibits the a/ß interferon system which is induced as host antiviral response. Moreover, it prevents the activation of double-stranded-RNA activated protein kinase (PKR) by binding to dsRNA and inhibits the maturation of GAS8 (gene of tumor suppressor). NS1 protein is utilized as a target for diagnostic of influenza viruses in infected animals. pUC57 carrying NS1 synthetic gene of influenza B was attempted to be transformed into Escherichia coli strain BL21(DE3). NS1B was extracted and attempted to be cloned into prokaryotic expression vectors pET-32b, pET-32a, pQE-81L and pQE-80L, respectively using restriction digestion enzymes (SacI, PstI and HindIII). Then, recombinant DNA was attempted to be transformed into Escherichia coli strains BL21(DE3) and DH5a.