Centre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB, Edifici CRAG, Campus UAB, Cerdanyola del Vallès, Barcelona, Spain.
Institute for Integrative Systems Biology (I2SysBio), Universitat de València-CSIC, Paterna, Valencia, Spain.
Methods Mol Biol. 2023;2686:495-508. doi: 10.1007/978-1-0716-3299-4_23.
Developmental processes in multicellular organisms depend on the proficiency of cells to orchestrate different gene expression programs. Over the past years, several studies of reproductive organ development have considered genomic analyses of transcription factors and global gene expression changes, modeling complex gene regulatory networks. Nevertheless, the dynamic view of developmental processes requires, as well, the study of the proteome in its expression, complexity, and relationship with the transcriptome. In this chapter, we describe a dual extraction method-for protein and RNA-for the characterization of genome expression at proteome level and its correlation to transcript expression data. We also present a shotgun proteomic procedure (LC-MS/MS) followed by a pipeline for the imputation of missing values in mass spectrometry results.
多细胞生物的发育过程依赖于细胞协调不同基因表达程序的能力。在过去的几年中,许多生殖器官发育的研究都考虑了转录因子的基因组分析和全局基因表达变化,以构建复杂的基因调控网络。然而,对发育过程的动态观察还需要研究蛋白质组在表达、复杂性及其与转录组的关系。在本章中,我们描述了一种双重提取方法,用于在蛋白质组水平上对基因组表达进行特征描述,以及其与转录组表达数据的相关性。我们还介绍了一种 shotgun 蛋白质组学方法(LC-MS/MS),以及随后用于对质谱结果中的缺失值进行推断的流水线。