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当前位置: 首页 > 文章 > 猪流行性腹泻病毒在稳定表达猪ACE2基因的IPEC-J2细胞中的增殖研究 动物医学进展 2020 (5) 38-43
Position: Home > Articles > Study on Proliferation of a PEDV Strain on IPEC-J2 Cells Stably Expressing Porcine ACE2 Progress in Veterinary Medicine 2020 (5) 38-43

猪流行性腹泻病毒在稳定表达猪ACE2基因的IPEC-J2细胞中的增殖研究

作  者：
沈海燕;康桦华;刘志成;张建峰;张春红
单  位：
广东省农业科学院动物卫生研究所/农业农村部兽药与诊断学科群广东科学观测实验站/广东省畜禽疫病防治研究重点实验室
关键词：
猪小肠上皮细胞;血管紧张素转换酶2;猪流行性腹泻病毒;增殖
摘  要：
为获得稳定表达猪源血管紧张素转换酶2(ACE2)的细胞株,用以增殖猪流行性腹泻病毒(PEDV)。构建了稳定表达猪源ACE2的猪小肠上皮细胞系(IPEC-J2),并对PEDV在该细胞上的增殖进行了分析。将重组表达质粒pCMV3-porcine-ACE2-GFP转染至IPEC-J2细胞后,经潮霉素B筛选和单克隆细胞培养,获得IPEC-J2-ACE2细胞系。通过荧光定量RT-PCR和Western blot对单克隆细胞中ACE2基因的mRNA转录和蛋白表达水平进行鉴定。以PEDV-WH株感染IPEC-J2-ACE2细胞,通过间接免疫荧光试验和荧光定量RT-PCR检测发现,PEDV-WH株在IPEC-J2-ACE2细胞中比在IPEC-J2细胞中的增殖能力高。
译  名：
Study on Proliferation of a PEDV Strain on IPEC-J2 Cells Stably Expressing Porcine ACE2
作  者：
SHEN Hai-yan;KANG Hua-hua;LIU Zhi-cheng;ZHANG Jian-feng;ZHANG Chun-hong;Institute of Animal Health,Guangdong Academy of Agricultural Sciences,Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province,Ministry of Agriculture,Key Laboratory of Livestock Disease Prevention of Guangdong Province;
关键词：
intestinal porcine epithelial cell line J2;;porcine angiotensin-converting enzyme 2;;Porcine epidemic diarrhea virus;;proliferation
摘  要：
In order to get a cell line susceptive to porcine epidemic diarrhea virus(PEDV)infection,we used eukaryotic expression of porcine angiotensin-converting enzyme 2(ACE2)in intestinal porcine epithelial cell line J2(IPEC-J2).And the replication of PEDV in IPEC-J2 was analysed.Firstly,the recombinant plasmid pCMV3-porcine-ACE2-GFP was transfected to IPEC-J2 cells.In order to obtain the monoclonal cell line IPEC-J2-ACE2,the transfected cells were selected with hygromycin B and using limited dilution method to get.Then,we used real-time PCR and Western blot methods to analyze the mRNA and protein expression levels of ACE2.The results displayed that porcine ACE2 protein was exactly and stably expressed in IPEC-J2-ACE2 cells.Moreover,we used PEDV-WH strain to infect IPEC-J2-ACE2 cells,then the results of indirect immunofluorescence assay and real-time PCR indicated that PEDV can replicated in IPEC-J2-ACE2 cells.Comparing to the control group that PEDV infected IPEC-J2 cells,the relative mRNA expression of PEDV N gene in PEDV infected IPEC-J2-ACE2 cells is higher than that in control group.It showed that PEDV is more susceptible to IPEC-J2-ACE2 cells.In conclusion,IPEC-J2-ACE2 cell has been successfully got,which is not only possibly useful for study the replication and vaccine of PEDV,but also lays the foundation for the research of pathogenic mechanism of PEDV.

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