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鸡传染性法氏囊病病毒VP2基因的克隆及序列分析

Cloning and Sequence of VP2 Gene of Chicken Infectious Bursal Disease Virus Ts and CJ-801 Srrain

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【作者】 刘小军; 陈楠; 陈永福;

【Author】 Liu Xiaojun; Chen Nan; Chen Yongfu(National Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100094)

【机构】 中国农业大学农业生物技术国家重点实验室!北京100094;

【摘要】 用幼鸡成纤维细胞繁殖鸡传染性法民囊病病毒(IBDV)天水(Ts)毒株和CJ-801毒株。用提纯的病毒颗粒提取基因组RNA。根据已报道的IBDV英国52/70株的序列设计引物,用反转录-聚合酶链式反应(RT-PCR)进行cDNA扩增,获得的两个1558bp的片段分别克隆于pGEM-T载体,亚克隆后进行序列分析,并与已报道的4株IBDV的VP2基因的核苷酸序列及推导的氨基酸序列进行了比较。结果表明,不同毒株间核苷酸同源性介于97.75%~99.68%之间,其中Ts与CJ-801以及Ts、CJ-801与CuI、PBG69间的同源性最高,为98.99%。不同毒株间推导的氨基酸同源性介于97.98%~98.99%之间。其中Ts与CJ-801以及Ts、CJ-801与CuI、PBG98间的同源性最高,为98.99%。Ts与CJ-801高变区内两个亲水区域与其它标准Ⅰ型毒株完全相同,而在致病毒株中保守的325~332位7肽区中,Ts和CJ-801的第5位丝氨酸突变为精氨酸,说明Ts与CJ-801为标准Ⅰ型弱毒株。

【Abstract】 The infectious bursal disease virus Ts strain and CJ-801 strain were propagated in 10-day-old specific-pathogenfree chicken embryo fibroblast Viral dsRNA was extracted and puritied. A Pair of primers was designed according to the segment A sequence of IBDV 52/70 strain rePOrted previously. Two separate cDNA fragmentS in length 1558hp were synthesize and amplified by RT-PCR. The PCR products were cloned intO PGEM-T vector and further subcloned into the vectors pGEM-7Zf. The compndive analysis indicated that the isolates are all closely relayed, ranging from 97J5 % to 99.68 % homologous. The Ts, CJ-801 and PBG98 isolates are the most identical with 99.68 % homology. Present sequence homology of the deduced afnino acids of the VP2 gene of the isolates ranging from 97.98 % to 98.99 %. The Ts, CJ-801, Cul, PBG98 isolates are the most identical with 98.99 % homology. In addition, the amino acids sequence of two hydrophilic regions in the variable region of VP2 was wholly conserved among the isolates, but a su bstitution from the third serine to arginine occurred in the heptCrpeptide which might be resPOnsible for the pathogenicity of IBDV strains in Ts and CJ-801 strains be standard serotype Ⅰstrain and have low pathogenicity.

【基金】 国家自然科学基金
  • 【文献出处】 农业生物技术学报 ,JOURNAL OF AGRICULTURAL BIOTECHNOLOGY , 编辑部邮箱 ,1999年02期
  • 【分类号】Q785
  • 【被引频次】7
  • 【下载频次】76
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