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中国农学通报

中国农学通报 ›› 2008, Vol. 24 ›› Issue (10): 13-16.

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

猪瘟病毒河南野毒株E2基因的克隆及序列分析

胡慧   

  • 收稿日期:2008-06-05 修回日期:1900-01-01 出版日期:2008-10-05 发布日期:2008-10-05

Cloning and sequence analysis of E2 gene of CSFV

  • Received:2008-06-05 Revised:1900-01-01 Online:2008-10-05 Published:2008-10-05

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摘要： 【研究目的】旨在了解猪瘟病毒流行毒株的变异情况，为防制猪瘟提供科学依据。【方法】根据GenBank上已发表的猪瘟石门株E2基因序列，设计合成了1对特异引物，应用RT-PCR技术对河南省一猪场采集的疑是猪瘟病料的猪瘟野毒E2基因进行扩增，将扩增的E2基因克隆于pGEM-T Easy载体，对其进行了序列测定及氨基酸序列推导，同时将其与HCLV株、C株、Alfort株和Brecsia株进行了同源性比较及遗传进化分析，并构建了CSFV的遗传发生树。【结果】通过PCR扩增得到与预期大小相等的产物，克隆测序验证是E2基因；所扩增的流行野毒与HCLV株、C株、Shimen株、Alfort株和Brecsia株核苷酸序列同源性分别为99.0%，96.5%，93.5%，94.4%和89.9%；氨基酸同源性分别97.8%，94.0%，92.6%，92.0% 和91.0%；所绘制的遗传发生树显示所测得流行野毒与C株、HCLV株关系较近。【结论】所测的流行株与与我国使用的C株疫苗毒核苷酸同源性很高，说明现行的猪瘟疫苗在一定程度上仍然可以起到保护作用。

Abstract: 【OBJECTIVE】To study the genetic variation of field virulent CSFV. 【METHOD】 The special primers were designed and synthesized according to the full E2 sequence of CSFV Shimen strain published in GenBank. The E2 gene of field virulent strain that was collected from Henan province was amplified by RT-PCR and cloned into pGEM-T vector. The nucleotide sequence was determined by sequencing and the amino acid sequence was deduced. 【RESULTS】The result showed that the full fragment of E2 gene was cloned. Compared with the corresponding region of HLCV, C, Shimen, Alfort and Brescia strain of CSFV, the nucleotide sequence homology was 99.0%, 96.5%, 93.5%, 94.4% and 89.9% respectively. And the amino acid homology was 97.8%, 94.0%, 92.6%, 92.0% and 91.0%, 87.0% respectively. The further study of the field strain was done by the phylogenetic tree. The filed sequenced strain has closer relation with HLCV strain compared with the Shimen, Alfort and Brescia strain. 【CONCLUSION】The result showed the isolated CSFV strain has high homology with the vaccine strain, suggesting that little mutation was occurred in the strain we cloned, and the current CSFV vaccine could provide whole protection for this virulent area in some extent.