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猪α干扰素多基因家族的特征分析

Characterization of the porcine alpha interferon multigene family.

作者信息

Cheng Gong, Chen Weizao, Li Zuofeng, Yan Weiyao, Zhao Xin, Xie Jun, Liu Mingqiu, Zhang Hao, Zhong Yang, Zheng Zhaoxin

机构信息

State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, 220 Handan Road, Shanghai 200433, PR China.

出版信息

Gene. 2006 Nov 1;382:28-38. doi: 10.1016/j.gene.2006.06.013. Epub 2006 Jun 29.

Abstract

The availability of data on the pig genome sequence prompted us to characterize the porcine IFN-alpha (PoIFN-alpha) multigene family. Fourteen functional PoIFN-alpha genes and two PoIFN-alpha pseudogenes were detected in the porcine genome. Multiple sequence alignment revealed a C-terminal deletion of eight residues in six subtypes. A phylogenetic tree of the porcine IFN-alpha gene family defined the evolutionary relationship of the various subtypes. In addition, analysis of the evolutionary rate and the effect of positive selection suggested that the C-terminal deletion is a strategy for preservation in the genome. Eight PoIFN-alpha subtypes were isolated from the porcine liver genome and expressed in BHK-21 cells line. We detected the level of transcription by real-time quantitative RT-PCR analysis. The antiviral activities of the products were determined by WISH cells/Vesicular Stomatitis Virus (VSV) and PK 15 cells/Pseudorabies Virus (PRV) respectively. We found the antiviral activities of intact PoIFN-alpha genes are approximately 2-50 times higher than those of the subtypes with C-terminal deletions in WISH cells and 15-55 times higher in PK 15 cells. There was no obvious difference between the subtypes with and without C-terminal deletion on acid susceptibility.

摘要

猪基因组序列数据的可得性促使我们对猪α干扰素(PoIFN-α)多基因家族进行特征分析。在猪基因组中检测到14个功能性PoIFN-α基因和2个PoIFN-α假基因。多序列比对显示六个亚型的C末端缺失八个残基。猪α干扰素基因家族的系统发育树确定了各种亚型的进化关系。此外,进化速率分析和正选择效应表明,C末端缺失是基因组中的一种保留策略。从猪肝基因组中分离出八个PoIFN-α亚型,并在BHK-21细胞系中表达。我们通过实时定量RT-PCR分析检测转录水平。分别通过WISH细胞/水疱性口炎病毒(VSV)和PK 15细胞/伪狂犬病病毒(PRV)测定产物的抗病毒活性。我们发现,在WISH细胞中,完整PoIFN-α基因的抗病毒活性比具有C末端缺失的亚型高约2至50倍,在PK 15细胞中高15至55倍。有无C末端缺失的亚型在酸敏感性方面没有明显差异。

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