1株山东猪瘟病毒流行毒株全基因组的克隆与序列分析  

Cloning and analysis of the whole genome sequence of epidemic strain of SD02 of CSFV from Shandong province

作　　者：吴旭锦[1] 朱小甫[1] 朱辉[2] 

机构地区：[1]咸阳职业技术学院生物科技系,陕西咸阳712000 [2]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2009年第11期7-12,18,共7页Journal of Northwest A&F University(Natural Science Edition)

摘　　要：【目的】对分离自山东的猪瘟病毒流行毒株SD02进行全基因组序列克隆分析,揭示SD02分子生物学特征,为猪瘟流行毒株分子流行病学研究积累资料。【方法】根据Genbank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12对引物,应用RT-PCR技术,从SD02毒株的细胞毒中成功扩增出了11段cDNA,将这11段cDNA与pMD18-T载体连接并进行克隆、测序,测得的序列经过拼接得到全基因组序列。将SD02与13株参考毒株(39、ALD、Alfort187、Brescia、CAP、Glentorf、GPE、GXWZ02、HCLV、LPC、Parderborn、Riems和Shimen)的核苷酸序列及开放阅读框架编码的氨基酸序列进行比较。【结果】拼接后SD02全长12300bp,SD02与13株参考毒株的核苷酸序列同源性为85.4%～99.1%;氨基酸序列同源性为92.5%～98.8%。SD02与我国Shimen经典强毒株核苷酸序列同源性高达99.1%,氨基酸序列同源性为98.8%。系统发生树分析表明,SD02与Shimen、HCLV、Riems、Glentorf、Alfort187、ALD等毒株同属于GroupⅠ。【结论】SD02毒株在遗传特性和抗原性上仍属于经典毒株。[Objective] Cloning and analysis of the whole genome sequence of epidemic strain of SD02 revealed the SD02 molecular biology features and accumulated research data for molecular epidemiology of classical swine fever epidemic strain virus. [Method] According to the Shimen and HCLV strains＇ whole genome sequence in Genbank, and references to other documents, 12 pairs of primers were synthesized using RT-PCR technology,11 cDNA were amplified from the SD02 strain successfully,and these 11 cDNA were comeeted with the pMD18-T vector,cloned and sequenced. Finally SD02 was compared with 13 reference strains of the nucleotide sequence and the amino acid sequences encoded from open reading frame. [Result] Spliced 11 cDNA acquired 12 300 bp length of the whole genome sequence,SD02 and 39 ,ALD,Alfort187,Brescia, CAP, Glentorf, GPE, GXWZ02, HCLV, LPC, Parderborn, Riems and Shimen nucleotide homology were 85.4%- 99.1%, amino acid homology 92.5 %-98.8%. SD02 and Shimen strain nucleotide homology 99.1% ,the amino acid homology 98.8%. The phylogenetic tree analysis showed that the SD02 and Shimen, HCLV, Riems, Glentorf, Alfort187, ALD, and other strains belonged to Group Ⅰ. [Conclusion] SD02 still belongs to classical strain in the genetic and antigen characteristics.

关 键 词：猪瘟病毒 全基因组 序列分析 SD02株 

分 类 号：S858.28[农业科学—临床兽医学]