›› 2017, Vol. 44 ›› Issue (3): 854-860.doi: 10.16431/j.cnki.1671-7236.2017.03.033

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Construction of Baculovirus Expression System and Antigenicity Analysis of Infectious Hematopoietic Necrosis Virus Glycoprotein

XIE San-lei1, WEN Shu-xiang1, LUO Lin2, MA Zhi-hong2, LI Gui-ping1, LI Tie-liang2, JIANG Na2, XING Wei2, SUN Hui-ling1   

  1. 1. Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;
    2. Beijing Fisheries Research Institute, Beijing 100068, China
  • Received:2016-07-18 Online:2017-03-20 Published:2017-03-21

Abstract:

To study the major structural protein glycoprotein (G) of infectious hematopoietic necrosis virus (IHNV), glycoprotein gene (1 380 bp) was amplified by RT-PCR from IHNV. In order to construct a recombinant plasmid pFB-LIC-Bse-G, G gene was cloned into the baculovirus vector pFB-LIC-Bse. Then, the constructed plasmid pFB-LIC-Bse-G was transformed into E.coli DH10Bac. The recombinant bacmids rBacmid-G was got, and then it was transfected to insect Sf9 cells,and the recombinant baculovirus that contained G gene was obtained. Western blotting analysis and indirect immunofluorescence assay (IFA) showed that the recombinant G protein could be recognized by histidine monoclonal antibody (Anti-His) and anti-IHNV antibody of mouse. The results indicated IHNV G protein had been expressed correctly in Sf9 cells. The study laid a foundation for further studying the G protein structure, function and immunological characteristics.

Key words: infectious hematopoietic necrosis virus; glycoprotein; recombinant baculovirus

CLC Number: