传染性法氏囊病病毒VP2-4-3 cDNA的分子克隆和序列分析  

Molecular Cloning and Sequence Analysis of VP2-4-3 cDNA of Infectious Bursal Disease Virus

作　　者：刘存仁[1] 梁志清[2] 

机构地区：[1]山东大学生命科学学院,济南250100 [2]香港大学动物学系

出　　处：《畜牧兽医学报》2002年第4期366-370,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：教育部留学回国人员科研启动基金资助

摘　　要：从法氏囊组织分离IBDV超强毒株HK46并提取基因组RNA。以RNA为模板进行反转录合成cDNA第一链。采用长PCR扩增技术获得VP2 4 3cDNA全长片段。将PCR产物克隆到 pcDNA3 1(+)载体 ,得到重组质粒 pPP1。对 pPP1插入片段全长序列进行了测序并对其序列进行了分析。结果表明 ,VP2 4 3cDNA阅读框架由30 39bp组成 ,可编码 10 12个氨基酸组成的前体多聚蛋白。经比较得知 ,HK46超强毒株VP2 4 3氨基酸序列与经典毒株间存在 19～ 2 8个氨基酸的差异 ;与Harbin强毒株相差 32个氨基酸 ;而与超强毒株OKYM和UK6 6 1分别相差 2和 6个氨基酸 ,且它们的VP2序列完全相同。在HK46超强毒株所特有的 9个氨基酸中 ,3个位于VP2可变区 ,显示超强毒株其抗原性存在着变异。A very virulent HK46 strain of infectious bursal disease virus(IBDV)was isolated from chicken bursa and the genomic RNA for virus was extracted.The cDNA first strand was synthesized by reverse transcription using IBDV genomic RNA as the template.A full length cDNA fragment coding for VP2-4-3 was obtained by long PCR amplification,which was cloned into pcDNA3.1(+)vector.The recombinant plasmid pPP1 was screened and identified.The nucleotide sequence of VP2-4-3 cDNA for HK46 strain was determined.The results showed that the open reading frame of VP2-4-3 cDNA consisted of 3039bp and the deduced amino acids were 1012.The amino acid sequence of HK46 strain was compared with those of other IBDV strains.The results indicated that there were 19-28 amino acid residues differences among VP2-4-3 of HK46 strain and other classical strains of IBDV.The VP2-4-3 of HK46 strain showed 32 amino acid differences from those of Harbin strain.But there were only 2 or 6 amino acid residues differences between HK46 and OKYM or UK661 strains.Moreover,VP2 amino acid sequence for HK46,OKYM and UK661 strains was identical.It has been found that there are 9 unique amino acids to HK46 strain.Three of them are in VP2 variable region.The results show vvIBDV strain has possibly variations for antigenecity.

关 键 词：VP2-4-3cDNA 分子克隆 序列分析 传染性法氏囊病病毒 病毒蛋白 

分 类 号：S852.659.4[农业科学—基础兽医学]