中国畜牧兽医

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不同毒力传染性法氏囊病病毒衣壳蛋白酵母双杂交诱饵载体的构建及鉴定

陈玉明,张礼洲,任宪刚,高立,秦立廷,高玉龙,王永强,高宏雷,王笑梅,祁小乐   

  1. (中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室,禽传染病研究室,黑龙江哈尔滨 150001)
  • 出版日期:2014-01-20 发布日期:2014-02-27
  • 通讯作者: 祁小乐(1980—),男,河南人,副研究员,主要从事动物传染病研究。E-mail:qxl@hvri.ac.cn 王笑梅(1962—),女,黑龙江人,研究员,主要从事动物传染病研究。E-mail:xmw@hvri.ac.cn
  • 作者简介:陈玉明(1988—),男,河北人,硕士生,研究方向:动物病毒学。
  • 基金资助:

    国家重点实验室基本科研业务费(SKLVBP201303);现代农业产业技术体系建设专项基金(nycytx-42-G3-01)。

Construction and Identification of Yeast Two-hybrid Bait Vectors of Capsid Protein of IBDV Different Virulence Strains

CHEN Yu-ming, ZHANG Li-zhou, REN Xian-gang, GAO Li, QIN Li-ting, GAO Yu-long, WANG Yong-qiang, GAO Hong-lei, WANG Xiao-mei, QI Xiao-le   

  1. (Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,Harbin 150001,China)
  • Online:2014-01-20 Published:2014-02-27

摘要: 传染性法氏囊病病毒(infectious bursal disease virus,IBDV)毒株Gx(超强毒株)、F9(中等毒力毒株)、Gt(弱毒株)遗传背景高度相似,但生物学性状差异显著。为研究不同毒力毒株与宿主细胞相互作用的分子细节,本研究将IBDV Gx、F9、Gt毒株的衣壳蛋白VP2基因克隆入pGBKT7载体,分别构建了诱饵载体pGBGxVP2、pGBF9VP2和pGBGtVP2。经Matchmaker Gold Yeast Two-hybrid System验证,结果显示所构建的3个诱饵载体均无自激活作用,对酵母细胞无毒性作用。本研究为利用酵母双杂交系统深入研究IBDV与宿主相互作用奠定了基础。

关键词: 传染性法氏囊病病毒; 衣壳蛋白; 酵母双杂交; 诱饵载体; 自激活作用; 毒性作用

Abstract: The infectious bursal disease virus (IBDV) strains,Gx (very virulent strain), F9 (intermediate strain) and Gt (attenuated strain) had highly similar genetic background, but their biological characteristics were significantly different. To investigate molecular details of the interaction between various IBDV and their host cells, the genes of VP2 capsid protein of Gx, F9 and Gt were cloned into vector pGBKT7, and the recombinant bait vectors pGBGxVP2, pGBF9VP2 and pGBGtVP2 were constructed. Then the self-activation and toxicity of the bait vectors were tested using the Matchmaker Gold Yeast Two-hybrid System. The results showed that three bait vectors all had no self-activations and toxicities to yeast cells. This research benefited to further investigate the interaction between IBDV and its host cell using the yeast two-hybrid system.

Key words: infectious bursal disease virus (IBDV); capsid protein; yeast two-hybrid; bait vector; self-activation; toxicity