羊口疮病毒 GDZC 株锚蛋白基因的克隆及序列分析

摘要

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 pro-teins 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 trans-membrane 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.%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。