IHNV核衣壳蛋白在杆状病毒表达系统中的表达及鉴定

doi:10.7668/hbnxb.2014.03.008

华北农学报 ›› 2014, Vol. 29 ›› Issue (3): 36-40. doi: 10.7668/hbnxb.2014.03.008

IHNV核衣壳蛋白在杆状病毒表达系统中的表达及鉴定

谢三磊^1,2, 王雯慧¹, 孙惠玲²

1. 甘肃农业大学动物医学院, 甘肃兰州 730070;
2. 北京市农林科学院畜牧兽医研究所, 畜禽疫病防控技术北京市重点实验室, 北京 100097

收稿日期:2014-03-19 出版日期:2014-06-28
通讯作者: 孙惠玲（1971-），女，甘肃武威人，副研究员，博士，主要从事动物传染病防控研究。
作者简介:谢三磊（1987-），男，河南鄢陵人，在读硕士，主要从事病原微生物分子生物学与免疫学研究。
基金资助:
北京市农林科学院科技创新基金（CXJJ201316）

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

XIE San-lei^1,2, WANG Wen-hui¹, SUN Hui-ling²

1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2. Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China

Received:2014-03-19 Published:2014-06-28

摘要/Abstract

摘要： 采用RT-PCR方法从传染性造血器官坏死病病毒(IHNV)中扩增RNA获得1 176 bp的核衣壳蛋白(N)基因，将其插入于pFB-LIC-Bse杆状病毒载体中，构建了重组质粒pFB-LIC-Bse-N，转化至感受态细胞DH10Bac中，获得重组杆粒rBacmid-N。再将其转染至Sf9昆虫细胞，获得重组杆状病毒。IFA和Western Blot分析表明，重组N蛋白可以被辣根过氧化物酶(HRP)标记的组氨酸单抗(anti-His)识别，表明，IHNV N基因在昆虫细胞Sf9中获得了正确表达。为深入研究N蛋白的功能和免疫学特性奠定了基础。

关键词: 传染性造血器官坏死病病毒, 核衣壳蛋白基因, 重组杆状病毒

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

中图分类号:

S432.4

谢三磊, 王雯慧, 孙惠玲. IHNV核衣壳蛋白在杆状病毒表达系统中的表达及鉴定[J]. 华北农学报, 2014, 29(3): 36-40. doi: 10.7668/hbnxb.2014.03.008.

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.

http://www.hbnxb.net/CN/Y2014/V29/I3/36

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