表达鸡传染性支气管炎病毒S1蛋白重组腺病毒的构建及免疫原性的初步分析  

Construction and lmmunogenicity Detection of Recombinant Adenovirus Expressing S1 Gene of Infections Bronchitis Virus

作　　者：宋菲菲[1,2] 康震 齐艳君 袁颖烁[1,2] 张守峰[2] 张乐萃[1] 扈荣良[2] 

机构地区：[1]青岛农业大学动物科技学院,青岛266109 [2]军事医学科学院军事兽医研究所,长春130122 [3]寿光六和肉鸡养殖有限公司,潍坊262700

出　　处：《畜牧兽医学报》2012年第9期1449-1454,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

摘　　要：拟构建表达鸡传染性支气管炎病毒(IBV)QS10株S1蛋白的重组复制缺陷型人5型腺病毒,并在本动物上进行初步的免疫学试验。通过RT-PCR获得S1基因,插入腺病毒表达系统穿梭质粒,构建重组穿梭质粒pac-Ad5CMV-S1,转染293AD细胞获得表达S1蛋白的重组腺病毒。PCR检测显示,S1基因已重组到腺病毒基因组;间接免疫荧光和Western blot检测证明该重组病毒在293AD细胞中真实表达了具有免疫反应性的IBV S1糖蛋白。重组病毒培养滴度可达到107 TCID50.mL-1。免疫接种SPF鸡后,通过ELISA检测,免疫鸡产生了针对IBV的特异性抗体。作者成功构建了表达IBV S1蛋白的重组腺病毒,该重组病毒可在SPF鸡体内诱导产生IBV特异性抗体。To construct human adenovirus type 5 vectored recombinant virus expressing S1 glyco- protein gene of avian infectious bronchitis virus （IBV）, S1 gene of IBV QS10 strain obtained through RT-PCR was inserted into shuttle plasmid of the adenoviral expression system, forming pacAd5 CMV-S1, as well as the backbone plasmid, which was linearized with Pac I and trans- fected into 293AD cells. Detection with PCR revealed that the recombinant virus was successfully packaged with a titer up to 10^7TCID50·mL^-1. Furthermore, The results of Western blotting and indirect immunofluorescence assay （IFA） indicated that the S1 gene was significantly expressed. Sera antibodies against IBV of the SPF chicken immunized with the recombinant virus were detec- ted by enzyme-linked immunosorbent assay （ELISA）. The recombinant adenovirus expressing S1 gene of IBV was successfully constructed with potent capacity to induce the specific antibodies in chickens and had the good immunogenicity, which laid the foundation for further development of recombinant avian infectious bronchitis vaccine.

关 键 词：鸡传染性支气管炎病毒 S1蛋白 人5型腺病毒载体 免疫原性 

分 类 号：S852.659.6[农业科学—基础兽医学]