›› 2011, Vol. 38 ›› Issue (11): 77-82.

• 生物技术 • 上一篇    下一篇

小熊猫源犬瘟热病毒全基因序列的克隆及序列分析

王磊1,2,3, 王照2,4, 李天松1,2, 刘玉秀1, 冯娜1,3, 王胜乐1,2, 许薇薇1,2, 姜雪1,2, 高玉伟1,3 ,王铁成1,3, 杨松涛1,3, 夏咸柱1,3   

  1. 1. 中国人民解放军军事医学科学院军事兽医研究所,吉林长春 130062;2. 吉林农业大学动物科技学院,吉林长春 130062;3. 吉林省人兽共患病预防与控制重点实验室,吉林长春 130062;4. 吉林省地方病第一防治研究所,吉林白城 137000
  • 收稿日期:1900-01-01 修回日期:2011-08-07 出版日期:2011-11-20 发布日期:2011-11-20
  • 通讯作者: 夏咸柱

Genomic Sequence Determination and Analysis of Lesser Panda Canine Distemper Virus

WANG Lei1,2,3, WANG Zhao2,4, LI Tian-song1,2, LIU Yu-xiu1, FENG Na1,3, WANG Sheng-le1,2, XU Wei-wei1,2, JIANG Xue1,2, GAO Yu-wei1,3,WANG Tie-cheng1,3, YANG Song-tao1,3   

  1. 1. Institute of Military Veterinary,Academy of Military Medical Science of PLA,Changchun 130062,China;2. College of Animal Science and Technology,Jilin Agricultural University,Changchun 130062,China;3. Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Changchun 130062,China;4. the First Institute for Control of Endemic Disease of Jilin Province,Baicheng 137000,China
  • Received:1900-01-01 Revised:2011-08-07 Online:2011-11-20 Published:2011-11-20

摘要： 本研究对军事兽医研究所病毒二室分离驯化后的小熊猫源犬瘟热病毒(canine distemper virus,CDV)基因组进行全序列测定,并对其基因组特征及H基因遗传稳定性进行比较分析。根据GenBank公布的犬瘟热病毒全长基因组序列设计合成17对特异性引物,以小熊猫源犬瘟热病毒总RNA为模板,RT-PCR进行分段扩增,并克隆到pEASY-Blunt Simple载体中,经测序、拼接获得全长cDNA序列。结果显示,小熊猫源犬瘟热病毒全基因序列与GenBank登录号分别为AF014953、AY445077、AY542312、AY466011、AY386316、AF164967、EU716337、EU726268、AY443350、AB474397、GU138403和AY649446的12个不同毒株全基因序列的同源性分别为86.6%、92.4%、92.5%、92.5%、94.3%、96.3%、95.9%、87.1%、94.5%、92.3%、87.4%和94.6%,与标准强毒株A75/17株(AF164967)的亲缘关系最近,全基因同源性达96.3%,但与疫苗株Onderstepoort(AF014953)亲缘关系相对较远,同源性为86.6%。小熊猫源犬瘟热病毒H基因与其他不同地区具有代表性的30株CDV进化树分析显示,小熊猫源犬瘟热病毒属于Asia Ⅰ型,H蛋白中309－311位氨基酸残基所形成的潜在糖基化位点,为疫苗株没有而野毒株所共有的,并且可能与病毒的免疫原性有关。因此,致弱的小熊猫源CDV在预防免疫的针对性上可能强于已有的疫苗株。

关键词: 犬瘟热病毒; 全基因测序; 序列分析

Abstract: The canine distemper virus (CDV) isoladed from lesser panda and domesticated by the laboratory of genome sequence determination,genetic characteristics and genetic stability of the H gene. According to full-length genome sequence of canine distemper virus published in GenBank, 17 pairs of specific primers were designed and synthesized to amplify full-length cDNA as template with total RNA of lesser panda canine distemper virus, using the method of RT-PCR, and cloned into pEASY-Blunt Simple vector for determination of full-length cDNA sequences.The results showed that the homology of whole genome sequence of lesser panda source canine distemper virus and 12 different strains AF014953, AY445077, AY542312, AY466011, AY386316, AF164967, EU716337, EU726268, AY443350, AB474397, GU138403 and AY649446 is 86.6%, 92.4%, 92.5%, 92.5%, 94.3%, 96.3%, 95.9%, 87.1%, 94.5%, 92.3%, 87.4% and 94.6%, respectively. It had the closest genetic relationship with the standard virulent strain A75/17, gene homology was 96.3%, but compared the vaccine strain Onderstepoort and it had distant genetic relationship, only 86.6% homology.Phylogenetic analysis of H gene of lesser panda source canine distemper virus and other thirty CDV published in GenBank in different districts showed that, canine distemper virus of lesser panda source belonged to the AsiaⅠ, the potential glycosylation sites of H protein gene were formed by amino acid from 309 to 311 of H protein, the wild strains had it common but the vaccine strain didn't, and may be associated with the virus immunogenic. Therefore, attenuated panda source CDV in the prevention of immune targeted may be better than the existing vaccines strain.

Key words: canine distemper virus; genomic sequence determination; sequence analysis

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