《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (12): 3085-3093.doi: 10.16431/j.cnki.1671-7236.2016.12.001

• 生物技术 •    下一篇

羊口疮病毒GDZC株锚蛋白基因的克隆及序列分析

葛士坤, 张凯照, 刘健新, 王晴楠, 于梦, 陶攀, 宁章勇   

  1. 华南农业大学兽医学院, 广州 510642
  • 收稿日期:2016-06-16 出版日期:2016-12-20 发布日期:2016-12-22
  • 通讯作者: 宁章勇 E-mail:ningzhyong@126.com
  • 作者简介:葛士坤(1990-),男,山东聊城人,硕士生,研究方向:兽医病理学,E-mail:geshikun2016@163.com
  • 基金资助:

    广东省科技计划项目(2016A020210079)

Cloning and Sequence Analysis of Ankyrin Genes from Orf Virus GDZC Strain

GE Shi-kun, ZHANG Kai-zhao, LIU Jian-xin, WANG Qing-nan, YU Meng, TAO Pan, NING Zhang-yong   

  1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China
  • Received:2016-06-16 Online:2016-12-20 Published:2016-12-22

摘要:

为研究羊口疮病毒(Orf virus,ORFV)编码的锚蛋白(ankyrin,ANK)基因在感染宿主中的免疫调节作用,本试验从ORFV GDZC株感染的细胞病毒液中提取基因组DNA,用特异性引物进行PCR扩增、克隆出5个ANKs基因,并进行了基因序列及其编码蛋白的生物信息学分析。结果显示,获得的GDZC株ORFV008、ORFV123、ORFV126、ORFV128、ORFV129基因分别编码516、525、497、501和516个氨基酸,与OV-SA00毒株的核苷酸同源性最高,分别为98.3%、98.7%、97.9%、97.3%、98.0%。ORFV编码的5个ANKs都含有ANK结构域和F-box结构域,是结构保守蛋白。蛋白跨膜分析表明,ORFV123和ORFV128不含潜在跨膜区,ORFV129、ORFV008和ORFV126蛋白分别存在1、2和3个潜在跨膜区,且均不含信号肽。本研究结果为深入研究ANK在ORFV致病过程中作用和免疫逃逸机制提供了基础数据。

关键词: 羊口疮病毒; 锚蛋白; 克隆; 序列分析

Abstract:

To investigate the role of ankyrin proteins encoding by Orf virus (ORFV) in the immune modulation to host infection, five ankyrin (ANK) genes of ORFV GDZC strain were amplified using specific primers by PCR amplification and cloning from the viral genome DNA which extracted from virus infection cells,and their sequences and coding protein structure bioinformatics analysis were performed. The results showed that ORFV008, ORFV123, ORFV126, ORFV128 and ORFV129 consisted of 516, 525, 497, 501 and 516 amino acids, respectively, which shared a nucleotide identity of 98.3%, 98.7%, 97.9%, 97.3% and 98.0% with those of ORFV OV-SA00 strain, respectively. All of five ANK proteins contained ANK and F-box structural domain that showed they were structure conservative proteins. The analysis of protein transmembrane showed that ORFV123 and ORFV128 had no potential transmembrane domains, while the ORFV129, ORFV008 and ORFV126 had one, two and three potential transmembrane domains, respectively. And these ANK proteins had no signal peptide. These results provided the basic data for further study of the ANK proteins in the pathogenic and immune evasion mechanisms of ORFV.

Key words: Orf virus; ANK protein; cloning; sequence analysis

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