Variation analysis of the VP1 and 3ABC genes from yellow cattle isolates persistently infected by Foot-and-mouth disease virus.

Shen Xiao-yan, Cong Guozheng, Chang Huiyun, Li Xiangtao, Xie Qing-ge
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Abstract

The potential relationship between the establishment of Foot-and-mouth disease virus (FMDV) persistent infection and gene variation was identified by investigating the variation of VP1 and 3ABC genes from yellow cattle persistent infection isolates. Five yellow cattle were inoculated on their tongue with 1.0×10 4 ID 50 /ml of FMDV O/Akesu/58 strain. After displaying clinical or subclinical signs, they probably became asymptomatic carriers. Oesophageal–pharyngeal fluids were collected monthly from the carriers with a probang and inoculated into a baby hamster kidney cell line (BHK-21); 12 FMDV isolates were obtained. The VP1 and 3ABC genes of the 12 isolates were then amplified by reverse-transcriptase polymerase chain reaction (RT-PCR). Cloning and sequencing revealed that the homology of the VP1 nucleotide and amino-acid sequence of all the isolates was above 98%, with no base deletion or insertion. When compared with the O/Akesu/58 FMDV strain, the homology of the VP1 nucleotide sequence of the isolates was only 85%, and that of the deduced amino-acid sequence only 90%.There were several nucleotide mutations in the VP1 gene of the isolates, including 16 consistent nucleotide mutations, with only two of them leading to a change in amino acid (I 56 →T, A 210 →E). Moreover, it was found that four nucleotide points and three amino-acid points had transversions among all isolates. The 3ABC gene had only 13 nucleotide transversions and five amino-acid mutations. It was presumed that persistent FMDV infection might have little connection with variation in the VP1 and 3ABC genes, and was probably related to other structural protein gene and key factors.
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持续感染口蹄疫病毒的黄牛分离株VP1和3ABC基因变异分析
通过对黄牛持续感染分离株VP1和3ABC基因变异的研究,确定了口蹄疫病毒(FMDV)持续感染的建立与基因变异之间的潜在关系。用1.0×10 4 id50 /ml的FMDV O/Akesu/58株在5头黄牛舌上接种。出现临床或亚临床症状后,可能成为无症状携带者。用probang每月从携带者身上收集食管咽液,并接种于幼鼠肾细胞系(BHK-21);分离得到12株口蹄疫病毒。利用逆转录聚合酶链反应(RT-PCR)扩增12株菌株的VP1和3ABC基因。克隆和测序结果表明,所有分离株VP1核苷酸和氨基酸序列同源性均在98%以上,无碱基缺失和插入。与O/Akesu/58 FMDV菌株的VP1核苷酸序列同源性仅为85%,推导出的氨基酸序列同源性仅为90%。分离株VP1基因存在多个核苷酸突变,包括16个一致的核苷酸突变,其中只有2个突变导致氨基酸的变化(I 56→T, a 210→E)。此外,发现4个核苷酸点和3个氨基酸点在所有分离株中都发生了翻转。3ABC基因只有13个核苷酸反转和5个氨基酸突变。推测FMDV持续感染可能与VP1和3ABC基因变异关系不大,可能与其他结构蛋白基因及关键因素有关。
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