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牛干扰素α1的特性:在毕赤酵母中的表达、生物学活性及理化特性

Characterization of bovine interferon α1: expression in yeast Pichia pastoris, biological activities, and physicochemical characteristics.

作者信息

Shao Jianwei, Cao Chong, Bao Jun, Liu Hongtao, Peng Tongquan, Gao Mingchun, Wang Junwei

机构信息

1 College of Veterinary Medicine, Northeast Agricultural University , Harbin, China .

出版信息

J Interferon Cytokine Res. 2015 Mar;35(3):168-75. doi: 10.1089/jir.2013.0139. Epub 2014 Oct 24.

Abstract

A bovine interferon α (BoIFNα) gene that included signal sequence was amplified from bovine liver genomic DNA. The gene was named BoIFN-α1 according to the position at which the encoded gene of the bovine IFN was located in the bovine genome. The sequence included a 23-amino-acid signal peptide and a 166-amino-acid mature peptide. The structural characteristics and phylogenetic relationships of the BoIFN-α1 gene were analyzed. A recombinant mature BoIFN-α1 (rBoIFN-α1) was expressed in the yeast Pichia pastoris. Physicochemical characteristics and antiviral activity were determined in vitro. Recombinant BoIFN-α1 was found to be highly sensitive to trypsin and stable at pH 2.0 or 65°C. It also exhibited antiviral activity, which was neutralized by a rabbit anti-rBoIFNα polyclonal antibody. This study revealed that rBoIFN-α1 has the typical characteristics of IFNα and can be used for both research and industrial application.

摘要

从牛肝脏基因组DNA中扩增出包含信号序列的牛干扰素α(BoIFNα)基因。根据牛干扰素编码基因在牛基因组中的位置,将该基因命名为BoIFN-α1。该序列包含一个23个氨基酸的信号肽和一个166个氨基酸的成熟肽。分析了BoIFN-α1基因的结构特征和系统发育关系。在毕赤酵母中表达了重组成熟BoIFN-α1(rBoIFN-α1)。在体外测定了其理化特性和抗病毒活性。发现重组BoIFN-α1对胰蛋白酶高度敏感,在pH 2.0或65°C下稳定。它还表现出抗病毒活性,该活性可被兔抗rBoIFNα多克隆抗体中和。本研究表明,rBoIFN-α1具有干扰素α的典型特征,可用于研究和工业应用。

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