O型口蹄疫病毒不同宿主适应毒株P1和3A基因的差异分析

发布时间：2018-01-20 16:54

本文关键词： O型口蹄疫病毒不同宿主适应毒株基因位点差异性　出处：《畜牧兽医学报》2017年05期 　论文类型：期刊论文

【摘要】：为了明确O型口蹄疫病毒不同宿主适应毒株P1和3A基因位点的差异性,研究其遗传变异趋势,找出不同宿主适应毒株在P1和3A基因水平上位点的变异情况,将O型口蹄疫病毒O/XJ/10-11株分别接种于牛、乳鼠、猪及BHK-21细胞,获得相应的宿主适应毒株,提取RNA,反转录并扩增P1和3A基因,目的片段产物经琼脂糖凝胶电泳回收,并将其克隆到pMD19-T载体上,筛选阳性菌落,测序并分析其基因序列。结果表明:牛、乳鼠、猪、BHK-21细胞四种宿主适应毒株,核苷酸和氨基酸序列均未发生缺失,主要抗原位点稳定;P1基因发生了部分变异,变异程度依次为VP1、VP2VP3VP4,VP4基因最为保守;3A基因较稳定,变异较少;不同宿主适应毒株的变异程度依次如下:猪适应毒株BHK-21细胞适应毒株牛适应毒株鼠适应毒株。O型口蹄疫病毒在不同宿主传代过程中造成VP1基因的变异,但主要抗原位点稳定;VP2基因的变异均位于抗原表位上;鼠适应毒株作为口蹄疫原始毒种的保存更为有利。
[Abstract]:In order to identify the difference of P1 and 3A gene loci in different hosts of foot-and-mouth disease virus type O (FMDV), the genetic variation trend of FMDV was studied. To find out the variation of P1 and 3A gene sites in different host adapted strains, O / XJ / 10-11 strain of foot-and-mouth disease virus (FMDV) was inoculated into bovine, mouse, pig and BHK-21 cells respectively. The corresponding host adapted strains were obtained, RNAs were extracted, P1 and 3A genes were amplified by reverse transcription. The products were recovered by agarose gel electrophoresis and cloned into pMD19-T vector. The positive strains were screened, sequenced and their gene sequences analyzed. The results showed that four host adapted strains of bovine, neonatal rat and pig BHK-21 cells had no deletion of nucleotide and amino acid sequences. The main antigen sites were stable; Partial variation of P1 gene occurred, and the degree of variation was that VP2VP3VP4VP4 gene was the most conserved. 3A gene is stable and has less variation. The degree of variation of different host adapted strains was as follows:. Porcine adaptive strain BHK-21 cell adapted strain bovine adaptive strain mouse adapted strain .O foot-and-mouth disease virus caused variation of VP1 gene during passage of different hosts. But the main antigenic sites were stable. The variation of VP2 gene was located on antigen epitope. It is more favorable for mice to adapt to the virus strain as the original strain of foot-and-mouth disease.
【作者单位】：新疆农业大学动物医学学院;天康生物股份有限公司动物用生物制品技术国家地方联合工程实验室;新疆畜牧科学院兽医研究所动物临床医学研究中心;
【基金】：新疆维吾尔自治区产学研联合培养研究生示范基地项目(xjaucxy-yjs-20152008) 新疆维吾尔自治区科研机构创新发展专项资金(2016D04008)
【分类号】：S852.65
【正文快照】： 口蹄疫病毒(foot-and-mouth disease virus,FMDV)属小RNA病毒科口疮病毒属,为无囊膜病毒,其基因组为单股正链RNA,全长约8.5kb[1-2]。P1基因编码的FMDV结构蛋白是构成病毒衣壳的基础,能诱导与感染性FMDV粒子相同的免疫反应。衣壳蛋白中VP1能诱导动物产生中和抗体,VP2具有抗原性

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