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禽脑脊髓炎病毒VP1基因的克隆与序列分析
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吕晓星1，鲁杏华1，朱忠武2，周辉煌3
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（1.湖南省兽医总站，湖南 长沙410007；2.湖南出入境检验检疫局，湖南 长沙410007；
3.湘阴县动物防疫检疫站，湖南 岳阳414600）
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摘要：
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为了揭示禽脑脊髓炎病毒（AEV）的分子特性，使用RT-PCR方法扩增出AEV SD和HB分离株和1株标准鸡胚适应株VR株的VP1基因，克隆到载体pBluescript SK(-)上，然后进行序列测定，并对这3株AEV的VP1基因与已发表的AEV活疫苗株1143株和强毒株L2Z株进行比较。结果，发现它们之间核苷酸和氨基酸的同源性分别为94.2%～99.9%和98.9%～100.0%。SD株和HB株与鸡胚适应毒VR株的VP1蛋白氨基酸序列完全相同。与1143株和L2Z株比较，VP1蛋白仅存在2～3个氨基酸的差异。结果表明，在不同AEV毒株中，VP1蛋白高度保守，存在极低的抗原性差异，不同AEV毒株的毒力和组织亲嗜性可能与VP1基因无关，VP1蛋白可作为防制AEV潜在的亚单位疫苗和诊断抗原。 |
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关键词：
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禽脑脊髓炎病毒（AEV）；VP1基因；克隆；序列测定 |
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中图分类号：S 852.659.6
文献标识码：
A 文章编号：1673-4696(2008)06-0470-05 |
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Cloning and sequence analysis of VP1 gene of avian
encephalomyelitis virus |
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L Xiao-xing1，LU Xing-hua1，ZHU
Zhong-wu2，ZHOU Hui-huang3 |
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(1.Hunan Provincial General Veterinary Station,Changsha 410007,China;2. Hunan Entry-Exit Inspection and
Quarantine Bureau,Changsha 410007,China;3.Animal Epidemic Prevention Department of Xiangyin County,
Yueyang 414600,China） |
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Abstract: To reveal the molecular characteristics of avian encephalomyelitis virus(AEV),VP1 gene sequences of AEV strains SD,HB and VR were determined.The VP1 genes of two AEV isolates and one standard egg-adapted strain VR were amplified by using RT-PCR,then cloned into the vector pBluescript SK(-) and sequenced.The sequences of VP1 genes of these three strains were compared with those of the live AEV vaccine strain 1143 and virulent strain L2Z.The results showed that the 94.2%-99.9% nucleic acid identities and 98.9%-100.0% amino acid similarities existed among the AEV strains examined so far.The two strains SD and HB had identical amino acid sequences with that of the strain VR.Furthermore,they had only 2-3 amino acid substitutions compared to the strains 1143 and L2Z.The results indicated that AEV VP1 was a highly conservative protein with much low antigenic variation among different strains,and the difference of virulent and tissue-tropism of different AEV strains might not be associated with the VP1 gene,which was used as a potential candidate for subunit vaccine and diagnostic antigen of AEV. |
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Key words: avian encephalomyelitis virus(AEV)；VP1 gene；cloning；sequencing |
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