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羽毛作为鸟类基因组研究中RNA的来源。

Feathers as a source of RNA for genomic studies in avian species.

作者信息

Jones Stephanie P, Kennedy Sean W

机构信息

Environment Canada, National Wildlife Research Centre, Carleton University, 1125 Colonel By Drive, Raven Road, Ottawa, ON, K1A 0H3, Canada,

出版信息

Ecotoxicology. 2015 Jan;24(1):55-60. doi: 10.1007/s10646-014-1354-z. Epub 2014 Sep 25.

Abstract

Dioxins and dioxin-like chemicals (DLCs) cause a suite of adverse effects in terrestrial species. Most of the adverse effects occur subsequent to binding to the aryl hydrocarbon receptor. Avian species vary in their sensitivity to the effects of DLCs and current research indicates that this is mediated by variations in the amino acid sequence within the ligand binding domain (LBD) of the aryl hydrocarbon receptor 1 (AHR1). Eighty-eight avian species have been classified into three broad categories of sensitivity, based on the amino acid variations within the AHR1 LBD: sensitive type 1 (Ile324_Ser380), moderately sensitive type 2 (Ile324_Ala380), and relatively insensitive type 3 (Val324_Ala380). Risk assessment of avian species can be complicated due to the variability in sensitivity among species. A predictive tool for selecting the priority species at a given site would have broad implications for the risk assessment community. We present a method for AHR1 genotyping using plucked feathers as a source of RNA. The method is extremely robust, requires minimal sample processing and handling, and eliminates the need for blood sampling or tissue collection from the species of interest. Using this method we were able to determine the amino acid sequence of the AHR LBD of three avian species: the chicken, the herring gull, and the zebra finch, and to categorize them based on the identity of amino acids at key sites within the LBD.

摘要

二噁英及二噁英类化学物质(DLCs)会对陆生物种产生一系列不利影响。大多数不利影响发生在与芳烃受体结合之后。鸟类物种对DLCs影响的敏感性各不相同,目前的研究表明,这是由芳烃受体1(AHR1)配体结合域(LBD)内氨基酸序列的变异介导的。根据AHR1 LBD内的氨基酸变异,88种鸟类已被分为三大类敏感性:敏感1型(Ile324_Ser380)、中度敏感2型(Ile324_Ala380)和相对不敏感3型(Val324_Ala380)。由于物种间敏感性的差异,鸟类物种的风险评估可能会很复杂。一种用于在给定地点选择优先物种的预测工具将对风险评估界产生广泛影响。我们提出了一种使用拔下的羽毛作为RNA来源进行AHR1基因分型的方法。该方法非常可靠,所需的样本处理和操作极少,并且无需从感兴趣的物种采集血液样本或组织样本。使用这种方法,我们能够确定三种鸟类(鸡、银鸥和斑胸草雀)AHR LBD的氨基酸序列,并根据LBD内关键位点的氨基酸身份对它们进行分类。

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