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一种新型牛干扰素-ε的分子克隆与特性分析

Molecular cloning and characterization of a novel bovine IFN-ε.

作者信息

Guo Yongli, Gao Mingchun, Bao Jun, Luo Xiuxin, Liu Ying, An Dong, Zhang Haili, Ma Bo, Wang Junwei

机构信息

Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang 150030, PR China.

Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang 150030, PR China.

出版信息

Gene. 2015 Mar 1;558(1):25-30. doi: 10.1016/j.gene.2014.12.031. Epub 2014 Dec 16.

Abstract

A bovine IFN-ε (BoIFN-ε) gene was amplified from bovine liver genomic DNA consisting of a 463bp partial 5'UTR, 582bp complete ORF and 171bp partial 3'UTR, which encodes a protein of 193 amino acids with a 21-amino acid signal peptide and shares 61 to 87% identity with other species IFN-ε. Then BoIFN-ε gene was characterized, and it can be transcribed in EBK cells at a high level after being infected by VSV. Recombinant proteins were expressed in Escherichia coli and the antiviral activity was determined in vitro, which revealed that bovine IFN-ε has less antiviral activity than bovine IFN-α. In addition, an immunofluorescence assay indicated that BoIFN-ε expressed in MDBK cells could be detected by polyclonal antibody against BoIFN-ε. Furthermore, the BoIFN-ε gene can be constitutively expressed in the liver, thymus, kidney, small intestine and testis, but not in the heart. This study revealed that BoIFN-ε has the typical characteristics of type I interferon and can be expressed constitutively in certain tissue, which not only can be a likely candidate for a novel, effective therapeutic agent, but also facilitate further research on the role of bovine IFN system.

摘要

从牛肝脏基因组DNA中扩增出牛干扰素ε(BoIFN-ε)基因,其由463bp的部分5'非翻译区(UTR)、582bp的完整开放阅读框(ORF)和171bp的部分3'UTR组成,编码一种含21个氨基酸信号肽的193个氨基酸的蛋白质,与其他物种的干扰素ε具有61%至87%的同一性。然后对BoIFN-ε基因进行了表征,在被水疱性口炎病毒(VSV)感染后,它能在EBK细胞中高水平转录。重组蛋白在大肠杆菌中表达,并在体外测定其抗病毒活性,结果表明牛干扰素ε的抗病毒活性低于牛干扰素α。此外,免疫荧光分析表明,在MDBK细胞中表达的BoIFN-ε可被抗BoIFN-ε的多克隆抗体检测到。此外,BoIFN-ε基因可在肝脏、胸腺、肾脏、小肠和睾丸中组成性表达,但在心脏中不表达。本研究表明,BoIFN-ε具有I型干扰素的典型特征,可在某些组织中组成性表达,这不仅可能成为一种新型有效治疗剂的候选物,还有助于进一步研究牛干扰素系统的作用。

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