稳定表达猪IRF7基因的PK-15细胞系建立及其抗病毒效果评价  

Establishment of a PK-15 cell line stably expressing porcine IRF7 gene and its effect on viral replication

作　　者：赵永祥[1] 李永涛[1] 赵军[1] 陈陆[1] 常洪涛[1] 王川庆[1] 

机构地区：[1]河南农业大学禽病研究所,河南郑州450001

出　　处：《中国兽医学报》2015年第12期1991-1996,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31272567);高校博士学科点专项科研基金资助项目(2011410511003);河南省高校科技创新团队资助项目(14IRTSTHN015)

摘　　要：为探讨猪IRF7基因对某些猪源病毒在PK-15细胞系中增值的影响,利用Piggy Bac真核转座子系统将猪源IRF7基因转染入PK-15细胞中,通过嘌呤霉素和绿色荧光进行双重筛选获得阳性转化细胞,再经克隆纯化得到单个阳性细胞克隆株。通过观察细胞荧光和荧光定量PCR鉴定,最终获得了稳定表达IRF7基因的PK-15细胞系。在Poly I:C诱导下,该细胞系能够显著上调IFN-β的表达。初步研究了该细胞系对病毒复制的影响,结果表明过表达IRF-7基因能够显著抑制猪伪狂犬病病毒(PRV)和猪水泡性口炎病毒(VSV)的复制。本研究为进一步探索猪IRF7基因的功能与作用机制提供有效工具。In order to study the effect of IRF7 gene on the replication of some porcine viruses,por- cine IRF7 gene was transfected into PK-15 cells using PiggyBac transposon system. The positive cells were screened by puromycin selection and fluorescence detection. A single clone of transfected PK-15 cells expressing porcine IRF7 was generated by several rounds of cloning and sub-cloning and identified by qRT-PCR assay and fluorescence detection. The transfected PK-15 cells stably expressing porcine IRF7 could upregulate the production of IFN-β with the stimulation of polyI：C and suppress the replication of porcine pseudorabies virus（PRV） and vesicular stomatitis virus （VSV） ,respectively. The PK-15 cell line stably expressing porcine IRF7 provides a useful tool for the study on the other functions of porcine IRF7 gene.

关 键 词：IRF-7基因 PiggyBac转座子系统 PK一15细胞系 抗病毒 

分 类 号：S852.4[农业科学—基础兽医学] S855.3[农业科学—兽医学]