传染性法氏囊病病毒超强毒GZ株VP2基因的克隆和序列分析

Cloning and Sequence Analysis of the VP2 Gene of GZ Strain of Very Virulent Infectious Bursal Disease Virus

根据已发表的传染性法氏囊病病毒 ( IBDV) 5 2 /70株基因组序列 ,设计并合成了 1对特异扩增 IBDVVP2基因的引物。以 IBDV超强毒 ( vv IBDV) GZ株基因组为模板 ,利用 RT-PCR技术扩增出了 1 .5 kb的c DNA产物 ,将 VP2基因克隆于 p UC1 1 9质粒上 ,得到重组 p UC1 1 9质粒。对 VP2基因全序列进行了测定。序列分析和聚类分析表明 ,GZ株与欧洲超强毒株 UK6 6 1非常相似 ,而与经典强毒株。

According to the published sequence of IBDV strain 52/70,a pair of primers that can amplify the cDNA of protective antigen VP2 gene was designed and synthesized.By RT PCR,a single DNA fragment of about 1 5 kb was obtained from GZ strain of vvIBDV.Then the VP2 cDNA was cloned into pUC119 at SmaⅠ site.The nucleotide sequence of the expected VP2 gene was determined by Sanger′s DNA sequencing method,and then the amino acid sequence was deduced.Both the nucleotide sequence and deduced amino acid sequence were compared with five published sequence of VP2 gene of IBDV strains.It showed that GZ strain is most closely related to the very virulent strain UK661 and OKYM,but different from other serotype I strains.