| 摘要: |
| [目的]对我国福建省流行的猪瘟病毒E0基因进行序列测定及分析.[方法]应用RT-PCR,从我国福建省送检的54份病料中扩增出14份猪瘟病毒E0基因,将其克隆到T载体上并测序.利用DNAstar分析软件,对所测定的14株流行毒株与参考毒株的相应基因片段进行同源性及序列分析.[结果]得到约801 bp的猪瘟病毒E0基因片段.序列分析表明,14株福建流行毒株可分为2个基因群,其中FJ216与ALD、Alfort187、Brescia、C HVRI,GPE、Glentorf、CAP、Shimen和HCLV等我国主要参考毒株属于基因1群,其他13株流行毒株和我国另一分离株GXWZ02属于基因2群.基因1群的FJ216与HCLV的核苷酸和氨基酸同源性分别为95.0%和94.3%,与Shimen的核苷酸和氨基酸同源性分别为99.4%和99.1%;而另外13株流行毒株之间核苷酸和氨基酸同源性分别为89.7%～99.9%和93.84%～99.6%,13株流行毒株与HCLV株的核苷酸和氨基酸同源性分别为81.8%～88.1%和86.8%～89.5%,与Shimen株的核苷酸和氨基酸同源性分别为83.4%～90.8%和88.6%～93.9%.得到的14株猪瘟病毒ED基因具有RNase活性的2个区域均高度保守,没有发生变异.[结论]福建省近期流行的猪瘟病毒以基因2群为优势毒株,而且大多数与HCLV、Shimen毒株之间存在较大差异. |
| 关键词: 猪瘟病毒 E0基因 基因克隆 序列分析 |
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| 基金项目:农业部中国猪病监测项目? |
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| Cloning and sequence analysis of genome E0 of CSFV isolated from Fujian |
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| Abstract: |
| 【Objective】 The study was to analyze the E0 gene sequence of classical swine fever virus(CSFV) in Fujian,China.【Method】 A total of 14 DNA fragments of E0 gene isolated from 54 pathological tissues,from various regions in Fujian were amplified by RT PCR,cloned,sequenced and compared with those of 10 reference strains Alfort187,ALD,Brescia,GPE,C HVRI,Shimen,GXWZ02,Glentorf,CAP and HCLV by using DNAstar software. 【Result】 The nucleotide sequence and the amino acid sequence of the 24 HCV strains shared an 81.8％-99.9％and an 86.8％-99.6％ homology respectively.The 24 HCV strains were divided into 2 major groups,HCV groupⅡbeing represented by FJ216,Shimen,HCLV and so on,and HCV group Ⅱ by the other HCV strains of Fujian province and GXWZ02.But the two motifs of Erns RNase activity，SLHGIWPG(or E) (29-36 sites) and EWNKHGWC (75-82 sites) were highly conserved, there being only one mutation from E to O in the domain EWNKHGWC in the 14 field isolates,and no mutation being found in the key amino acid residue H located in the 30 and 79 sites.【Conclusion】 Of the E0 gene of the 14 field isolates,HCV was found to be complex,diverse and stable. |
| Key words: CSFV E0 gene gene cloning sequence analysis |
