发布: 2020年08月20日第10卷第16期 DOI: 10.21769/BioProtoc.3727 浏览次数: 2817
评审: Arvind PandayFabian den BraveFernando A Gonzales-Zubiate
Abstract
The yeast Saccharomyces cerevisiae has been perceived over decades as a highly valuable model organism for the investigation of ion homeostasis. Indeed, many of the genes and biological systems that function in yeast ion homeostasis are conserved throughout unicellular eukaryotes to humans. In this context, measurement of the yeast cellular ionic content provides information regarding yeast response to gene deletion or exposure to chemicals for instance. We propose here a protocol that we tested for the analysis of 12 elements (Ba2+, Ca2+, Cd2+, Co2+, Cu2+, Fe2+, K+, Mg2+, Mn2+, Na+, Ni2+, Zn2+) in yeast using Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES). This technique enables determination of the cellular content of numerous ions from one biological sample.
Keywords: Ion content (离子含量)Background
Yeast has been extensively used to study the ionic response after gene deletion, modification of the extracellular environment, or heterologous protein production for example. In this context, having methods to monitor the ionic status of yeast cells is of high interest. ICP-AES uses high-energy plasma from an inert gas like argon to burn analytes very rapidly. The color that is emitted from the analyte is indicative of the elements present, and the intensity of the spectral signal is indicative of the concentration of these elements. Compared with other techniques (for example spectrophotometric methods, atomic absorption spectrometry and atomic fluorescence spectrometry), ICP-AES is a multi-ion analysis method involving simple and fast procedure with relatively low detection limits (Dahlquist and Knoll, 1978). The method described here applies to concentration determination of Ba2+, Ca2+, Cd2+, Co2+, Cu2+, Fe2+, K+, Mg2+, Mn2+, Na+, Ni2+, and Zn2+, with the possibility of measurement of numerous cellular ionic concentrations from one 100 ml yeast culture (Eide et al., 2005; Thines et al., 2018). While this protocol has been tested in our laboratory for these twelve ions, this protocol could most likely be transferred to other elements. Besides, although described here for whole-cell measurement of the yeast ionic content, this method could be transferred to other cell types or to isolated organelles by fractionation on sucrose gradient for instance.
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© 2020 The Authors; exclusive licensee Bio-protocol LLC.
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分类
微生物学 > 微生物生物化学 > 其它化合物
细胞生物学 > 基于细胞的分析方法 > 离子分析
生物化学 > 其它化合物 > 离子
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