表达鸡γ-干扰素重组鸡传染性支气管炎病毒的构建及鉴定  

Construction and identification of recombination infectious bronchitis virus expressing interferon-γ

作　　者：姜逸[1] 唐梦君[1] 程旭[1] 沈欣悦[1] 李建梅[1] 尤素兰[1] 赵宝华[1] 戴亚斌[1] 周生[1] 

机构地区：[1]中国农业科学院家禽研究所,江苏扬州225003

出　　处：《中国预防兽医学报》2015年第3期167-171,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31101832);江苏省科技支撑计划(BE2012369)

摘　　要：为研究鸡传染性支气管炎病毒(IBV)作为载体表达外源基因的可行性,本研究以IBV疫苗株H120为病毒载体,在其非结构蛋白5a编码区上游插入鸡γ-干扰素(Ch IFN-γ)基因,通过反向遗传操作技术构建重组病毒。经RT-PCR和测序鉴定表明拯救获得重组病毒(r H120-c IFNγ/5a)。生物学特性研究结果表明,r H120-c IFNγ/5a感染鸡胚后能够引起特征性病变,但与其亲本病毒株H120相比,病毒的毒价及其在鸡胚中的复制能力有所下降。将r H120-c IFNγ/5a在鸡胚中连续传代,采用RT-PCR进行检测,结果表明Ch IFN-γ基因在病毒传至第9代时仍保持稳定,至第12代Ch IFN-γ基因出现部分或完全丢失现象。本研究结果为进一步研制以IBV为载体的新型基因重组疫苗奠定了基础。To investigate the feasibility of avian infectious bronchitis virus （IBV） as a vector for expressing exogenous gene, we used vaccine strain H120 of IBV as a vector to express the chicken IFN-y （ChlFN-y） gene. Then, the ChlFN-y gene was amplified by RT-PCR and inserted into the upstream of 5a gene by reverse genetic techniques. Rescued recombinant virus （rH120-clFNy/5a） expressing ChlFN-y gene was rescued and identified by RT-PCR and sequencing. The results of biological characteristics showed that rH120-clFNy/5a was still able to lead to chicken embryo dwarf. However, compared with the wild-type strain H120, the viral titer and replication rate of recombinant virus were reduced relatively. In addition, the rH120-clFNy/5a remained stable after the ninth passages in SPF embryonated eggs for continuous passages, but ChlFN-y gene in the recombinant virus lost partly or completely by passaged in chick embryo more than twelve passages. These results laid a foundation for further development of new recombinant IBV vaccine.

关 键 词：传染性支气管炎病毒 干扰素-Γ 反向遗传技术 重组病毒 

分 类 号：S852.65[农业科学—基础兽医学]