鸡传染性法氏囊病毒Ts、OH毒株、鱼传染性胰脏坏死病毒DRT毒株的同源性分析  

The Homology Analysis Among Infectious Bursal Disease Vrius Ts Strain、OH Strain and Infectious Pancreatic Necrosis Virus DRT Strain

作　　者：陈立栋[1] 林爱星[1] 周胜花[1] 刘小军[1] 周忠孝 

机构地区：[1]中国农业大学农业生物技术国家重点实验室,北京100094

出　　处：《农业生物技术学报》2000年第3期257-261,共5页Journal of Agricultural Biotechnology

基　　金：中国农业大学农业生物技术国家重点实验室开放课题基金

摘　　要：用幼鸡成纤维细胞繁殖鸡传染性法氏囊病病毒（IBDV）Ts毒株，病毒颗粒经提纯后提取基因组dsRNA。根据已报道的加拿大OH株序列设计引物，用RT-PCR进行 cDNA扩增，获得976 bp、774 bp和997 bp的三个部分重叠的片段，分别克隆于pGEM-Teasy载体，利用SP6，T7特异引物及PCR引物进行序列分析,并连接为一个完整的大片段。结果表明，所克隆片段为2 665 bp的IBDV VP1基因的大开放读框。序列分析表明：Ts株与OH株的核苷酸序列同源性为91 %；与DRT毒株的氨基酸序列同源性为42.5 ％。尽管Ts株与DRT株的整体同源性很低，但在VP1蛋白的中部仍发现有高同源区段。在两种病毒中均发现与GTP结合蛋白、依赖于RNA的RNA聚合酶（RdRp）相关的保守序列。然而，与单链RNA病毒不同的是：作为RdRp家族特征的G-D-D序列在两种病毒中都未发现。Infectious bursal disease virus Ts strain was propagated in 1 0-day-old specific-pathogen-free chicken embryo fibroblast.The dsRNA w as extracted and purified.Three pairs of primers were designed accordi ng to the segment B sequence of IBDV OH strain reported previously.Thr ee separate but overlapping cDNA fragments in length of 976 bp、774 bp and 997 bp were amplified by RT-PCR.The PCR products were cloned into pGEM-T easy vector.Special primers SP6、T7 and PCR primers were used to sequence.Sequencing showed that the larger ORF of IBDV Ts strain se gment B in length of 2 665 bp was cloned. The nucleotide sequence anal ysis indicated that the homology between Ts and OH Strain is 91 %.The comparative analysis of deduced amino acid sequence between IBDV Ts st rain and IPNV DRT strain indicated that the homology is only 42.5 %,De spite the low overall homology between the IPNV and IBDV VP1 Proteins, homologous regions were detected within the central protein of the pro teins.Consensus sequences associated with GTP-binding proteins and RdR ps were also detected in VP1.However,unlike the RdRps associated with single-stranded plus RNA viruses,the birnavirus RdRp lacks the Gly-Asp -Asp motif characteristic of this enzyme family.

关 键 词：鸡传染性法氏囊病毒 鱼传染性胰脏坏死病毒 

分 类 号：S858.312.6[农业科学—临床兽医学] S941.41[农业科学—兽医学]