甲型H1N1流感病毒NS1蛋白核仁定位的研究.pdfVIP

2012,20(3): 12-16 Chinese Journal of Animal Infectious Diseases H1N1 NS1 430070 2009 H1N1 NS1 RT-PCR NS1 PEGX-KG KG-NS1BL21IPTG GST NS1 Western blot NS1 NS1-219NS1- 230NS1-2373 3 NS1 NS1 H1N1 NS1 S853.659. 5: S852.43 A 1674-6422(2012)03-0012-05 LOCALIZATION OF NS1 PROTEIN OF H1N1 INFLUENZA A VIRUS IN NUCLEI ZHAO Zong-zheng, TU Jia-gang, ZHANG Wen-ting, HU Yong, ZHOU Hong-bo, JIN Mei-lin (State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China) Abstract: In order to investigate the distribution of NS1 protein of H1N1 Influenza A virus of 2009 in nuclei, the NS1 gene was amplifi ed in RT-PCR and cloned into PEGX-KG vector. The resulting plasmid KG-NS1 was then transformed into E.coli BL-21 and the recombinant protein NS1 was expressed with induction of IPTG. NS1 protein was purifi ed using GST affi nity chromatography and injected into rabbits for the preparation of anti-NS1 polyclonal antibodies. The production of rabbit anti-NS1 antibodies was determined in Western blotting. Then, the nucleic localization of NS1 protein with three different lengths (219, 230 and 237 amino acids) was examined in indirect immunofl uorescence using rabbit antibodies. The NS1 protein with three different lengths accumulated in the nucleus and cytoplasm but not in the nucleolus. The results showed that truncation of NS1 protein did not affect its nucleolus localization. Key words: H1N1 Infl uenza A virus; NS1; polyclonal antibody; nuclear localization