鸡传染性支气管炎病毒地方分离株S1基因克隆和基因型鉴定  

GENOTYPES IDENTIFICATION AND MOLECULAR CLONING OF S1 GENES OF AVIAN INFECTIOUS BRONCHITIS VIRUS ISOLATED IN CHINA

作　　者：祁贤[1] 叶向阳[1] 张婉华[1] 朱永军[1] 步志高[2] 刘思国[2] 

机构地区：[1]上海市农业科学院畜牧兽医研究所，上海 201106 [2]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，哈尔滨 150001

出　　处：《上海农业学报》2000年第4期52-56,共5页Acta Agriculturae Shanghai

基　　金：国家攀登计划B类项目"85-44-01-44"资助

摘　　要：利用IBV全长S1基因特异性引物 ,以纯化的 3个标准株M4 1、H52 、T和 6个地方分离株 (QD、ZZ、GZ、TJ、DL和YC)的基因组RNA为模板 ,用RT PCR方法扩增出预期大小约 1.73kbcDNA片段 ,经BAMHI和HindIII酶切插入到质粒 pUC18中 ,并在大肠杆菌JM 83中实现了克隆。对 6个分离株的S1基因PCR产物进行HaeIIIRFLP分析 ,表明QD、TJ属于M4 1基因型 ,ZZ和GZ与T基因型一致 ;而DL、YC表现为不同的基因型 ,这一结果与血清中和实验结果基本吻合 ,证实我国IBV流行株已发生基因变异。Based on the published IBV genomic sequences, a pair of primers were designed and synthesized for specific amplification of S1 glycoprotein genes of 3 reference strains M 41,H 52,T and six isolated strains(QD,ZZ,GZ,TJ,DL and YC).1.7kb cDNA fragments predicted were amplified by RT-PCR,using purified genomic RNA of IBV as templetes. Digested with BamhI/HindIII, cDNA of the S1 genes cDNA were inserted in BamhI/HindIII site of pUC18 plasmid to construct recombinant cloned in complement E.coli JM83 strain.The cDNA of S1 genes were digested,and the RFLP patterns indicated that QD and ZZ belong to M 41 genotypes, GZ and TJ belong to T genotype, and DL and YC are different from the reference strains. The results were consistent with the previously serotype identification by neutralisation test.

关 键 词：鸡传染性支气管炎病毒 S1基因 RFLP 基因型鉴定 

分 类 号：S858.315.3[农业科学—临床兽医学]