Influenza A virus (IAV) subtypes are globally responsible for seasonal pandemics. Subtypes of IAV virus can beclassified because of differences surface antigenic protein. The non structural protein1 (NS1) of IAV is the only protein that encoded by IAV and contributing role in the pathogensis. NS1 is a virulence factors duringviral replication in the host. NS1 of IAV is known to be multifunctional protein which enhances the pathogenicity of the virus duringt he viral life cycle. The current study is aimed to investigate NS1 of IAV subtype H1N1, H7N7, H4N6 and H5N1, which were successfully cloned in to the pcDNA 3.1 vector. Restriction analysis with enzymes BamH1,Xba1 and Kpn1 and sequencing showed that NS1 gene was successfully cloned. My recombinant proteins have been used by Dr. Broniaczyk to successfully express in cell culture.