鸡传染性支气管炎病毒国内分离D971株5a、5b及核蛋白基因的分子特征  

Cloning and Sequence Analysis of 5a、5b and Nucleocapsid Genes ofInfectious Bronchitis Virus D971 Isolated from China

作　　者：刘玉芬[1,2] 荣骏弓[1] 刘胜旺[1] 孔宪刚[1] 刘丽玲[1] 刘宝全[1,2] 

机构地区：[1]中国农业科学院,哈尔滨兽医研究所,兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]东北农业大学动物医学院

出　　处：《中国预防兽医学报》2003年第2期81-87,共7页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省政府博士后科研启动金资助项目(LRB_KY01045)

摘　　要：本研究参照已发表的鸡传染性支气管炎病毒 (Infectiousbronchitisvirus ,IBV)基因序列 ,设计合成了一对引物 ,经反转录_聚合酶链式反应 (ReverseTranscription_PolymeraseChainReaction ,RT_PCR)技术扩增得到了IBV国内分离株D971的 5a、5b与核蛋白 (Nucleocapsid ,N)基因 ,片段长约 1.7kb ,并将该基因进行了克隆和序列测定。与国内外部分已发表毒株比较表明 :D9715a基因与Beau、CU_T2、D14 6 6、DE0 72及SD/97/0 2毒株的 5a基因核苷酸序列同源性在 85 .9%～ 93.9%之间 ,氨基酸序列同源性为 78.5 %～ 93.8% ;D9715b基因与上述毒株的 5b基因核苷酸序列同源性在 91.6 %～ 96 .8%之间 ,氨基酸序列同源性为 87.8%～ 93.9% ,比 5a基因保守 ;N基因与Beau、CU_T2、Ark99、M4 1、H5 2、VicS、SD/97/0 2、X、HB、DB及ZJ971毒株的核苷酸序列同源性为 87.0 %～ 91.5 % ,氨基酸序列同源性为 89.7%～ 93.6 %。对文献报道的N基因三个保守位置同源性比较发现 ,D971N基因第 198～ 2 6 4位核苷酸及其推导的氨基酸与上述 11株毒株的相应区域同源性分别为 86 .6 %～ 92 .5 %和 90 .5 %～ 10 0 % ;第 5 4 3～ 70 2位核苷酸及其对应的氨基酸序列同源性分别为 79.4 %～ 90 .6 %和 83.3%～ 96 .3% ;第 993～ 112According to the published IBV structural gene sequence, a pair of specific primers were designed and synthesized. A fragment of 1.7Kb 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 the predicted amino acid were analysed with several published IBV strains. The results showed that the nucleotide sequence identity of 5a gene between D971 and Beau?CU_T2?D1466?DE072?SD/97/02 strains is from 85.9% to 93.9% and amino acid identity is from 78.5% to 93.8%, the nucleotide sequence identity of 5b gene between D971 and the above five strains is from 91.6% to 96.8% and amino acid identity is from 87.8% to 93.9%, the nucleotide sequence identity of N gene between D971 and Beau?CU_T2?Ark99?M41?H52 ?VicS?SD/97/02?X?HB?DB?ZJ971 is from 87.0% to 91.5% and amino acid identity is from 89.7% to 93.6%. The nucleotide sequence identity is from 86.6% to 92.5% in position 198_264 of N gene between D971 and the eleven IBV strains and amino acid identity is from 90.5% to 100%, the nucleotide sequence identity is from 79.4% to 90.6% in position 543_702 and amino acid identity is from 83.3% to 96.3%, the nucleotide sequence identity is from 87.1% to 96.0% in position 993_1122 and amino acid identity is 75.0%_95.0%. It was indicated that there were high sequence identity between D971 and SD/97/02 in 5a?5b gene and N gene in position 198_264. In addition, D971and ZJ971 shared high sequence identity in position 543_702 of N gene. Interestingly, in position 993_1122 of N gene, D971and H52.D971 showed high sequence identity. The results showed that D971 strain has unique molecular characterization.

关 键 词：传染性支气管炎病毒 5a、5b与N基因 分子特征 

分 类 号：S858.312.6[农业科学—临床兽医学] Q78[农业科学—兽医学]