Screening of Proteins Interacting with Nonstructural 1 Protein of H5N1 Avian Influenza Virus from T7-phage Display Library

Avian influenza virus(AIV) nonstructural 1 (NS1) gene was amplified by real-time polymerse chain reaction(RT-PCR) and inserted into pET28a, then transformed into E. coli BL21(DE3) competent cell. With the induction of isopropyl-beta-D-thiogalactoside(IPTG) and the purification of Ni-NTA column, we finally obtained purified NS1 protein. T7-phage display system was used to screen the proteins that interacted with NS1 from lung cell cDNA library. The selected positive clones were identified by DNA sequencing and analyzed by BLAST program in Gene-Bank. Two proteins were obtained as NS1 binding proteins, Homo sapiens nucleolar and coiled-body phosphoprotein 1(NOLC1) and Homo sapiens similar to colon cancer-associated antigen. By co-immunoprecipitation and other methods, Homo sapiens NOLC1 was found to interact with the NS1 protein, the results would provide the basis for further studying biological function of NS1 protein.