摘要
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.
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.
作者
ZHU Chun-yu1,2, SUN Ting-ting2, ZHAO Jian1,2, WANG Ning1,2, ZHENG Fang-liang1, AI Hai-xin1, ZHU Jun-feng1,2, WANG Xiao-ying3, ZHU Ying4, WU Jian-guo4 and LIU Hong-sheng1,2 1. Key Laboratory of Animal Resource and Epidemic Disease Prevention of Liaoning Province, 2. Research Center for Computer Simulating and Information Processing of Bio-macromolecules of Liaoning Province, Academy of Science, Liaoning University, Shenyang 110036, P. R. China
3. Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, P. R. China
4. State Key Laboratory of Virology, Wuhan University, Wuhan 430072, P. R. China
基金
Supported by the National Natural Science Foundation of China(No.30671852)
the Open Research Fund Program of the State Key Laboratory of Virology of China(Nos.2010009, 2007007)
the Research Fund of the Key Laboratory of Department of Education of Liaoning Province, China(No.2009S043)
