鸡传染性支气管炎病毒免疫原基因cDNA的克隆和序列分析  

CLONING AND SEQUENCING OF cDNA OF THE AVIAN INFECTIOUS BRONCHITIS VIRUS IMMUNOGEN GENE

作　　者：张玉根[1,2] 郭玉璞[1,2] 陈永福 

机构地区：[1]北京农业大学动物医学院 [2]北京农业大学农业生物技术国家重点实验室

出　　处：《病毒学报》1998年第1期54-59,共6页Chinese Journal of Virology

摘　　要：从含有鸡传染性支气管炎病毒（Ｉｎｆｅｃｔｉｏｕｓｂｒｏｎｃｈｉｔｉｓｖｉｒｕｓ，ＩＢＶ）的鸡胚尿囊液中快速提取病毒ＲＮＡ，得到约２３ｋｂ全长ＩＢＶＲＮＡ。运用反转录－聚合酶链式反应（ＲＴ－ＰＣＲ）得到１．７２ｋｂ编码全长ＩＢＶ免疫原糖蛋白Ｓ１的基因片段。进一步将此基因片段插入到质粒ｐＵＣ１９中，进行全长片段的序列分析。结果表明，ＩＢＶ北京株和Ｂｅａｕｄｅｔｅ株的核酸同源性为９７．８％，与Ｍ４１株的核酸同源性达９８．５％。Infectious Bronchitis Virus(IBV) Beijing strain RNA extracted from the allantoic fluid of inoculated embryonated eggs was converted to cDNA by reverse transcription with 3′-primer.Polymerase chain reaction was performed with two primers which span the S 1 spike region.Amplified product of 1.7kb was subjected to HaeⅢ digestion and the fragments obtained were the same as expected.The PCR product was ligated to pUC19 plasmid vector,and its nucleotide sequence was determined by the dideoxy-mediated chain termination method.A comparison of the nucleotide sequence with that of Beaudette and M41 strains showed that the homology was 97% and 98% respectively.The Beijing strain was identified as the Massachusetets serotype. This study may be helpful in rapid monitoring of disease incidence,serotyping of IBV and the development of IBV nucleic acid probes and genetically-engineered IBV vaccines.

关 键 词：IBV 免疫原 分子克隆 序列 鸡 传染性支气管炎 

分 类 号：S852.65[农业科学—基础兽医学] S858.316.3[农业科学—兽医学]