摘要
为了对分离到的猪瘟流行毒株SXYL2006进行全基因组序列克隆分析,揭示其遗传变异特征,为猪瘟分子流行病学研究提供素材,为猪瘟防控提供理论参考,根据Gen Bank上发表的古典猪瘟Shimen毒株及HCLV株全基因序列,参考有关文献合成了12对引物,应用RT-PCR技术,从陕西省猪瘟流行毒株SXYL2006中成功扩增出了11段c DNA,测序后拼接得到全长12 295 bp全基因序列,提交Gen Bank获得登录号GQ122383。将SXYL2006与参考毒株进行比较,结果表明SXYL2006与毒株39、ALD、Alfort187、Brescia、CAP、Glentorf、GPE、GXWZ02、HCLV、LPC、Paderborn、Riems和Shimen核苷酸序列同源性分别为88.2%、85.5%、85.4%、85.0%、85.1%、85.0%、85.2%、95.6%、84.4%、83.8%、94.8%、84.7%和85.1%,氨基酸序列同源性分别为94.2%、92.9%、92.9%、92.9%、92.1%、92.0%、92.5%、97.4%、91.6%、90.6%、97.5%、92.0%和92.5%。系统发生树分析结果表明SXYL2006与GXWZ02在进化上距离最近,亲缘关系最为密切,两者与德国流行毒株Paderborn同处于基因Ⅱ群。与HCLV株相比,SXYL2006株E0蛋白中,具有Rnase活性的2个主要氨基酸序列没有发生任何变异;在E2蛋白中,与免疫逃逸相关的5个位点有3个发生变异;已发现的26个T细胞表位中,有14个表位27个氨基酸位点变异。SXYL2006在遗传特性和抗原性上与经典强毒株Shimen、疫苗毒株HCLV差异较大。
To clone and analyze the complete genome sequence of classical swine fever virus strain SXYL2006 and reveal the genetic variation and molecular epidemiology of classical swine fever,eleven c DNA segments were amplified from SXYL2006 by RT-PCR with 12 pairs of primers synthesized according to the genome sequences of classical swine fever strains Shimen and HCLV published in Gen Bank. After sequencing and splicing,a full-length 12 295-bp genome sequence was obtained（ Gen Bank accession number GQ122383）. Compared with reference strains 39,ALD,Alfort187,Brescia,CAP,Glentorf,GPE,GXWZ02,HCLV,LPC,Parderborn,Riems and Shimen,the nucleotide of SXYL 2006 shared homologies of 88. 2%,85. 5%,85. 4%,85. 0%,85. 1%,85. 0%,85. 2%,95. 6%,84. 4%,83. 8%,94. 8%,84. 7% and85. 1%,respectively, and amino acid sequence shared94. 2%, 92. 9%, 92. 9%, 92. 9%, 92. 1%, 92. 0%,92. 5%,97. 4%,91. 6%,90. 6%,97. 5%,92. 0% and92. 5% homologies, respectively. Phylogenetic tree analysis showed that SXYL2006 and GXWZ02 were the closest evolutionarily,and the two strains and German virulent strain Paderborn were classified into gene group Ⅱ,compared with HCLV strains. There was no variation in the two major amino acid sequences with Rnase activity in protein E0 encoded by SXYL2006. Three out of five sites related to immune escape in protein E2 mutated. There were 27 amino acid mutated sites in 14 out of 26 T cell epitopes. SXYL2006 differed from strains Shimen and HCLV in genetic characteristics and antigenicity.
出处
《江苏农业学报》
CSCD
北大核心
2016年第1期133-141,共9页
Jiangsu Journal of Agricultural Sciences
基金
陕西省科学技术研究发展计划项目(2014K02-06-03)
咸阳市科学技术研究计划项目(2014K02-21)
