鸡传染性法氏囊病超强毒Gx及其致弱株基因组B节段的克隆和序列分析  

Cloning and sequencing of segment B of very virul Gx and attenuate strain of infectious bursal disease virus

作　　者：包晓玮[1,2] 张厚双[1] 高宏雷[1] 付朝阳[1] 彭志伟[1] 单文鲁[3] 王笑梅[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所 [2]新疆农业大学动物医学系,新疆乌鲁木齐830000 [3]新疆农业大学动物医学系

出　　处：《中国预防兽医学报》2005年第2期85-90,共6页Chinese Journal of Preventive Veterinary Medicine

摘　　要：用蛋白酶K法分离提取了鸡传染性法氏囊病病毒强毒株Gx及其致弱株Gt的病毒核酸dsRNA ,应用随机引物将RNA反转录成cDNA ,以此为模板用长距离一步法扩增出全长基因B节段 ,将其克隆入PMD18_T载体 ,进行了测序 ,并用DNAStar软件进行序列分析。测序结果表明 ,克隆的Gx株B节段全长为 2 82 7bp ,与超强毒参考株UK6 6 1的同源性为 88 2 % ,与强毒参考株Harbin_1株的同源性达 96 3% ;克隆的Gt株B节段全长为 2 82 7bp ,与弱毒参考株P2的同源性达 99 5 %。而Gx株与Gt株B节段的核苷酸的同源性只有 89 8% ;vvIBDV_Gx。The double stranded genomic RNA of Gx&Gt strain of infectious bursal disease were extracted by proteins K digestion based approach from chicken embryo fibroblast(CEF) adapted virus strain.By the methods of reverse transcription,followed by the Long_accurate polymerase chain reaction (LA_PCR) in a single step amplification of single stranded cDNA,and cloning of full_length cDNA of segment B of IBDV_Gx7Gt ,resulted in the synthesis of 2 827 bp of full_length Larger segment B.The amplified cDNA fragments were directly cloned into T Vector and analysed by DNAStar software.As a results,the sequence of Gx compare with the published standard V_virulent stains UK661 strain,the homology of the nucleotide was 88.2 %,the sequence of Gx compare with the published standard virulent stains Harbin_1,the homology of the nucleotide was 96.3 %;the Gt strain was 99.5 %,compare with the published standard attenuate strain P2 the homology of the Gx strain was only 89.8 %,compare with the Gt strain;That attainiog the full length of segment B of IBDV_Gx and IBDV_Gt enhance the necessary availability to research on the construction of infectious clones of infectious bursal disease virus.

关 键 词：鸡传染性法氏囊病病毒 基因组B节段 克隆和序列分析 

分 类 号：S852.65[农业科学—基础兽医学] Q785[农业科学—兽医学]