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Cloning and sequence analysis of H and F genes of a canine distemper vi- rus isolated from fox

LU Pin HU Chuanwei YANG Dubao XIE Zhijing   

  1. A nimal Science and Technology Institute,Shandong Agricultural University,T ai′an 271018,Chi na
  • Online:2008-10-09 Published:2008-07-07

狐源犬瘟热病毒H 基因和F 基因的克隆与序列分析

吕品 胡传伟 杨笃宝 谢之景   

  1. 山东农业大学动物科技学院,泰安271018

Abstract: Based on the H and F genes of canine distemper virus isolates reported in GenBank (CDV 5804(AY386315)、CDV 01-2689(AY649446)、CDV A75-17(AF164967)、CDV 00-2601(AY443350)、CDV 98-2645(AY445077)、CDV 98-2654(AY466011)and CDV Onderstepoort(AF378705)),two pairs of primers were designed. The H and F genes of CDV-FOX-TA were amplified by RT-PCR,and were cloned into pMD18-T vector,which were sequenced and analyzed. The H gene of CDV-FOX-TA contained a ORF including 1 824 bp,and had the lower identity with the CDV vaccine strains (Onderstpoort and Convac ),only 89.2% and 90.6% ,respectively. However,it had 98.6% identity with CDV A75-17, and 98.7% identity with CDV LP which was isolated from lesser panda. The ORF of CDV-FOX-TA F gene was 1 889 bp. The major difference among various CDV strains was in the signal peptide domain,but the RRQRR,the thirteen serine residues, the four protential asparagines glycosylation sites and the two hydrophobic regions were highly conserved. According to phylogenetic analysis,it showed that CDV-FOX-TA,CDV A75 - 17 and CDV LP had the nearer inherited distance.

Key words: Canine distemper virus Fox F gene H gene

摘要: 根据GenBank 发表的犬瘟热病毒(CDV)核酸序列[CDV5804(AY386315)、CDV01-2689(AY649446)、CDV A75-17 (AF164967)、CDV 00-2601 (AY443350)、CDV 98-2645(AY445077)、CDV 98-2654 (AY466011)和CDV Onderstepoort(AF378705)],分别设计合成扩增CDV H 基因和F 基因的引物。采用RT-PCR 技术扩增狐源CDV 泰安株(CDV-FOX-TA)H 基因和F 基因,分别将H 基因与F 基因克隆进pMD18-T 载体,进行序列分析。结果表明,CDV-FOX-TA 株H 基因有1 个ORF,全长1 824 bp,编码607 个氨基酸,与CDV Onderstpoort、CDV Convac 的同源性分别为89.2% 和90.6% ,与CDV A75-17 的同源性为98.6% ,与CDV LP 的同源性为98.7% 。CDV-FOX-TA F 基因有1 个ORF,全长1 889 bp,编码662 个氨基酸,CDV F 蛋白信号肽区域易发生突变,但F0蛋白裂解点(RRQRR)、13 个半胱氨酸残基、4个潜在的天冬氨酰糖基化位点和2个疏水区都高度保守。CDV H基因和F 基因系统发生分析表明,CDV-FOX-TA 与CDV A75-17 和CDV LP 亲源关系较为密切。

关键词: 狐狸 犬瘟热病毒 H基因 F基因