摘要
参照已发表的鸡传染性支气管炎病毒 (infectious bronchitis virus,IBV)基因序列 ,设计合成了 1对引物 ,经 RT-PCR扩增得到了 IBV国内分离株 D971约 1.3kb的片段 ,其中包含 3a、3b、E及 M蛋白基因的完整 ORF,并将该片段进行了克隆和序列测定。与国内外部分 IBV毒株序列比较表明 :D9713a基因与 CU- T2、D14 6 6、DE0 72、M4 1、GA/99和 UK/6 6的 3a基因核苷酸序列同源性在 83%～ 89%之间 ,氨基酸序列同源性为 77%～ 91% ,其中与 CU- T2毒株的同源性最低 (分别为 83%和 77% ) ;3b基因与上述毒株的对应核苷酸序列同源性在 85 %～ 95 %之间 ,氨基酸序列同源性为 72 %～ 96 % ,与 CU- T2的同源性最高 (分别为 95 %和 96 % ) ;E蛋白基因与 CU- T2、D14 6 6、DE0 72、M4 1、Ark99、H5 2、Gray、GA/99和 UK/6 6的核苷酸序列同源性在 81%～ 86 %之间 ,氨基酸序列同源性为 74 %～ 80 %。不同毒株推导的 E蛋白 C末端表现不同特征 ,其中 D971E蛋白 C末端比 CU- T2的对应区多 9个氨基酸残基 ,而比本试验选用的其他参考毒株的相应区域短 5～ 7个氨基酸残基。M基因与 CU - T2、DE0 72、Gray、M4 1、H12 0、H5 2、SD/97/0 2、L X4及D4 1的 M基因核苷酸序列同源性在 86 %～ 91%之间 ,氨基酸序列同源性为 85 %～ 95 % ,其?
According to the published IBV gene sequence,a pair of specific primer was designed and synthesized.A fragment of 1.3 kb including mRNA_3 and mRNA_4 cDNA of IBV strain isolated from China was amplified by reverse transcription-polymerase chain reaction(RT-PCR) method and cloned into pMD18-T vector.The fragment was sequenced and the sequences of nucleotide and predicted amino acid were analyzed with several published IBV strains.The results showed that the nucleotide sequence identity of ORF3a gene between D971 and CU-T2,D1466,DE072,M41,GA/99,UK/66 is from 83% to 89% and amino acid identity is from 77% to 91%,the nucleotide sequence identity of ORF 3b gene between D971 and the above six strains is from 85% to 95% and amino acid identity is from 72% to 96%.The nucleotide sequence identity of E gene between D971 and CU-T2,D1466,DE072,M41,Ark99,H52,GA/99,UK/66,Gray is from 82% to 86% and amino acid identity is from 74% to 80%.The nucleotide sequence identity of M gene between D971 and CU-T2,DE072,Gray,M41,H120,H52,SD/97/02,LX4,D41 is from 86% to 91% and amino acid identity is from 85% to 95%.It was also indicated that there were high sequence identity between D971 and SD/97/02 in position 1-19 at N-terminal of M amino acid.In addition,D971 and SD/97/02 shared 100% identity in position 20-67 and 101-182 of M amino acid.Interestingly,D971 had 100% identity with H52 in position 184-219 and 97% identity with H120 in position 101-182 and 184-219 of M protein.Mutation also exists in position 220-222 and 224 at C terminal sequence of M protein of D971 as compared with other reference strains.All these data above showed that IBV D971 shared more characteristics with SD/97/02 isolated from China than other reference strains.In addition,the relationship between D971 and the two vaccine IBV strains,namely H52 and H120,was also found in M gene and its functional regions.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2004年第2期122-125,共4页
Chinese Journal of Veterinary Science
基金
黑龙江省政府博士后科研启动资金资助项目 (LRB-KY0 10 45 )
