Expression and Identification of Nucleocapsid Protein of IHNV in Baculovirus Expression System

doi:10.7668/hbnxb.2014.03.008

Abstract

Abstract: To study the major structural nucleoprotein (N) of infectious hematopoietic necrosis virus(IHNV) by baculovirus systems, in this study nucleoprotein (N) gene(1 176 bp) was amplified by RT-PCR from infectious hematopoietic necrosis virus (IHNV) and inserted into the baculovirus vector pFB-LIC-Bse to construct a recombinant plasmid pFB-LIC-Bse-N.The constructed recombinant transposition plasmid pFB-LIC-Bse-N was transformed to competent E.coli DH10Bac to get recombinant bacmids rBacmid-N.The obtained recombinant bacmid rBacmid-N was transfected to insect Sf9 cells, and the recombinant baculovirus that contained N gene was obtained.IFA and Western Blot analysis showed that the recombinant N protein can be recognized by horseradish peroxidase (HRP) labeled histidine monoclonal antibody (anti-His) which indicated IHNV N gene has been successfully expressed in Sf9 cells infected with recombinant baculovirus.The study laid a foundation of further study on N protein structure, function and immunological characteristics.

Key words: Infectious hematopoietic necrosis virus, N gene, Recombinant baculovirus

CLC Number:

S432.4

XIE San-lei, WANG Wen-hui, SUN Hui-ling. Expression and Identification of Nucleocapsid Protein of IHNV in Baculovirus Expression System[J]. journal1, 2014, 29(3): 36-40. doi: 10.7668/hbnxb.2014.03.008.

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