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猪干扰素诱导跨膜蛋白抑制JEV和EMCV的复制
发表时间：2016-11-15 浏览量：1786 下载量：451
| 全部作者： | 王星晨，盖新娜，郭 鑫 |
| 作者单位： | 中国农业大学动物医学院 |
| 摘 要： | 为研究猪干扰素诱导跨膜蛋白（interferon-induced transmembrane family members，IFITM）家族的全基因序列及其抗病毒性能，经实时荧光定量PCR（real-time PCR，RT-PCR）和cDNA末端快速扩增（rapid amplification of cDNA ends，RACE）技术扩增、克隆并测序，共获得3个猪IFITM家族蛋白。利用pCMV-Myc载体构建重组质粒pCMV-IFITM1/2/3，通过Western blotting检测方法确认重组质粒在BHK-21细胞系中表达情况，通过RT-PCR和噬斑方法检测日本乙型脑炎病毒（Japanese encephalitis virus，JEV）和脑心肌炎病毒（encephalomyocarditis virus，EMCV）在IFITM过量表达下的复制情况。结果表明，猪IFITM家族与人IFITM家族结构相似，但3端亲水区多出13个氨基酸。猪IFITM家族的3个蛋白均可以抑制JEV和EMCV在体外易感细胞中的复制，其中以IFITM3抗病毒效果最显著。本研究结果丰富了IFITM抗病毒谱，为干扰素刺激基因在人畜共患病防治中的应用奠定了基础。 |
| 关 键 词： | 兽医免疫学；干扰素诱导跨膜蛋白；抗病毒；日本乙型脑炎病毒；脑心肌炎病毒 |
| Title： | Porcine interferon-induced transmembrane proteins: inhibitors of JEV and EMCV replication |
| Author： | WANG Xingchen, GE Xinna, GUO Xin |
| Organization： | College of Veterinary Medicine, China Agricultural University |
| Abstract： | In order to study the full gene sequences and antiviral properties of porcine interferon- induced transmembrane family members (IFITM), three of them were amplified using real-time PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) and cloned into the vector pCMV-Myc separately, and recombinant plasmid pCMV-IFITM1/2/3 were obtained. Overexpression of these three proteins in BHK-21 cell lines was identified by Western blotting. The replication abilities of both Japanese encephalitis virus (JEV) and encephalomyocarditis virus (EMCV) when IFITM was overexpressed were detected using RT-PCR and plaques. Our data reveal that the amino acid sequences of porcine IFITM family members are quite similar to that of human IFITM proteins, except for the additional 13 amino acids in the 3 terminal hydrophilic region. All of the three porcine IFITM family members inhibit the replication of JEV and EMCV in vitro, and the IFITM3 protein shows the strongest antiviral activities. These results have enriched the antiviral spectrum of IFITM proteins and laid the foundation for the application of interferon-stimulated genes in zoonosis prevention. |
| Key words： | veterinary immunology; interferon-induced transmembrane protein; antiviral; Japanese encephalitis virus; encephalomyocarditis virus |
| 发表期数： | 2016年11月第21期 |
| 引用格式： | 王星晨，盖新娜，郭 鑫. 猪干扰素诱导跨膜蛋白抑制JEV和EMCV的复制[J]. 中国科技论文在线精品论文，2016，9（21）：2246-2251. |
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