稳定表达鸡氨肽酶N的MDCK细胞系构建及其对传染性支气管炎病毒感染的影响  

Construction of chicken aminopeptidase N stabilized expressed MDCK cell line and its effect on infectious bronchitis virus infection

作　　者：李广兴[1] 郭德启[1] 王雪[1] 葛旭影 任玉东[2] 任晓峰[1] 黄小丹[1] 张瑞莉[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]东北农业大学电气与信息学院,哈尔滨150030

出　　处：《东北农业大学学报》2015年第10期63-67,共5页Journal of Northeast Agricultural University

基　　金：国家自然科学基金项目(31172295;31272569)

摘　　要：为研究鸡氨肽酶N(Chicken aminopeptidase N,c APN)在鸡传染性支气管炎病毒(IBV)感染细胞过程中作用,构建稳定表达c APN的MDCK细胞系。将重组质粒pc DNA3.1-c APN通过脂质体转染到MDCK细胞中并利用G418筛选,克隆纯化获得稳定表达c APN的MDCK细胞系。通过RT-PCR和间接免疫荧光检测表明c APN在MDCK细胞中持续稳定表达。IBV可以感染稳定表达c APN的MDCK细胞并连续传代,该细胞系接种IBV阳性样品24 h即可产生典型细胞融合病变,而对照MDCK细胞接种后没有细胞病变出现。结果表明c APN在介导IBV感染宿主细胞过程中发挥重要作用。To investigate the biological role of chicken aminopeptidase N（c APN） in the process of avian infectious bronchitis virus（IBV） infection of host cel s, MDCK cel s were transfected with c APN plasmid of pc DNA3.1-c APN, and the positive cel s was cloned and screened under the drug selection of G418. The stabilized expression of c APN positive cel line was confirmed by RT-PCR and indirect immunofluorescence assay. Virus infection experiment demonstrated that IBV could infect and serial propagated in this cel line, and typical cel pathogenic effect of cel fusion could also be observed 24 h post inoculation, while no changes appeared in the control cel s. It implies the critical biological role of c APN during IBV infection.

关 键 词：鸡传染性支气管炎病毒 鸡氨肽酶N 稳定表达细胞系 病毒感染 

分 类 号：S852.23[农业科学—基础兽医学]