Sáenz Johan S, Seifert Jana
Institute of Animal Science, University of Hohenheim, Stuttgart, Germany.
HoLMiR-Hohenheim Center for Livestock Microbiome Research, University of Hohenheim, Stuttgart, Germany.
Methods Mol Biol. 2024;2820:49-56. doi: 10.1007/978-1-0716-3910-8_6.
The development of high throughput methods has enabled the study of hundreds of samples and metaproteomics is not the exception. However, the study of thousands of proteins of different organisms represents different challenges from the protein extraction to the bioinformatic analysis. Here, the sample preparation, protein extraction and protein purification for livestock microbiome research throughout metaproteomics are described. These methods are essential because the quality of the final protein pool depends on them. For that reason, the following workflow is a combination of different chemical and physical methods that intend an initial separation of the microbial organisms from the host cells and other organic materials, as well as the extraction of high concentrate pure samples.
高通量方法的发展使得对数百个样本的研究成为可能，宏蛋白质组学也不例外。然而，研究不同生物体的数千种蛋白质，从蛋白质提取到生物信息学分析，都面临着不同的挑战。在此，描述了贯穿宏蛋白质组学的家畜微生物组研究的样品制备、蛋白质提取和蛋白质纯化。这些方法至关重要，因为最终蛋白质库的质量取决于它们。因此，以下工作流程是不同化学和物理方法的组合，旨在初步从宿主细胞和其他有机材料中分离微生物，以及提取高浓缩的纯样品。
