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 reac- tion(RT-PCR) and inserted into pET28a, then transformed into E. coli BL21(DE3) competent cell. With the induction of isopropyl-β-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 li- brary. 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 me- thods, Homo sapiens NOLC1 was found to interact with the NS1 protein, the results would provide the basis for fur- ther studying biological function of NS1 protein. Keywords T7-phage display; Nonstructural 1(NS1) protein; Interacting protein Article ID 1005-9040(2012)-01-103-05