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与 Concholepas concholepas 病原体反应相关的泛素结合酶 E2 样基因:SNP 鉴定与转录表达。

Ubiquitin-conjugating enzyme E2-like gene associated to pathogen response in Concholepas concholepas: SNP identification and transcription expression.

机构信息

Laboratorio de Biotecnología y Genómica Acuícola, Departamento de Oceanografía, Facultad de Ciencias Naturales y Oceanográficas, Universidad de Concepción, P.O. Box 160-C, Concepción, Chile.

出版信息

Fish Shellfish Immunol. 2012 Oct;33(4):1065-8. doi: 10.1016/j.fsi.2012.08.024. Epub 2012 Sep 3.

Abstract

Ubiquitin-conjugated E2 enzyme (UBE2) is one of the main components of the proteasome degradation cascade. Previous studies have shown an increase of expression levels in individuals challenged to some pathogen organism such as virus and bacteria. The study was to characterize the immune response of UBE2 gene in the gastropod Concholepas concholepas through expression analysis and single nucleotide polymorphisms (SNP) discovery. Hence, UBE2 was identified from a cDNA library by 454 pyrosequencing, while SNP identification and validation were performed using De novo assembly and high resolution melting analysis. Challenge trials with Vibrio anguillarum was carried out to evaluate the relative transcript abundance of UBE2 gene from two to thirty-three hours post-treatment. The results showed a partial UBE2 sequence of 889 base pair (bp) with a partial coding region of 291 bp. SNP variation (A/C) was observed at the 546th position. Individuals challenged by V. anguillarum showed an overexpression of the UBE2 gene, the expression being significantly higher in homozygous individuals (AA) than (CC) or heterozygous individuals (A/C). This study contributes useful information relating to the UBE2 gene and its association with innate immune response in marine invertebrates.

摘要

泛素缀合 E2 酶 (UBE2) 是蛋白酶体降解级联的主要组成部分之一。之前的研究表明,在受到某些病原体(如病毒和细菌)挑战的个体中,UBE2 的表达水平会增加。本研究通过表达分析和单核苷酸多态性 (SNP) 发现,来描述腹足纲动物海兔(Concholepas concholepas)UBE2 基因的免疫反应。因此,通过 454 焦磷酸测序从 cDNA 文库中鉴定出 UBE2,而 SNP 的鉴定和验证则是通过从头组装和高分辨率熔解分析来完成的。用鳗弧菌进行攻毒试验,以评估处理后 2 至 33 小时 UBE2 基因的相对转录丰度。结果显示,有一段 889 个碱基对 (bp) 的 UBE2 部分序列,其部分编码区为 291bp。在第 546 位观察到 SNP 变异 (A/C)。受到鳗弧菌攻击的个体表现出 UBE2 基因的过表达,纯合个体 (AA) 的表达明显高于杂合个体 (A/C)。本研究为海洋无脊椎动物的 UBE2 基因及其与先天免疫反应的关联提供了有用的信息。

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