Joseph Biju, Frosch Matthias, Schoen Christoph, Schubert-Unkmeir Alexandra
Institute for Hygiene and Microbiology, Würzburg, Germany.
Methods Mol Biol. 2012;799:267-93. doi: 10.1007/978-1-61779-346-2_17.
Infectious disease research has been revolutionized by two recent developments in the field of genome biology: (1) the sequencing of the human genome as well as many pathogen genomes and (2) the development of high-throughput technologies including microarray technology, proteomics, and metabolomics. Microarray studies enable a deeper understanding of the genetic evolution of pathogens and investigation of the determinants of pathogenicity on a whole-genome scale. Host studies, in turn, allow for an unprecedented holistic appreciation of the complexities of host cell responses at the molecular level. In combination, host-pathogen studies allow global analysis of gene expression in the infecting bacterium as well as in the infected host cell during pathogenesis, providing a comprehensive picture of the intricacies of pathogen-host interactions. In this chapter, we briefly explain the principles underlying DNA microarrays including the major points to consider when planning and analyzing microarray experiments and we describe in detail their practical application, using the interaction of Neisseria meningitidis with human endothelial or epithelial cells as examples.
(1)人类基因组以及许多病原体基因组的测序;(2)包括微阵列技术、蛋白质组学和代谢组学在内的高通量技术的发展。微阵列研究有助于更深入地了解病原体的遗传进化,并在全基因组范围内研究致病性的决定因素。反过来,宿主研究能够以前所未有的方式从分子水平全面认识宿主细胞反应的复杂性。综合起来,宿主 - 病原体研究能够对发病过程中感染细菌以及被感染宿主细胞中的基因表达进行全局分析,全面呈现病原体与宿主相互作用的错综复杂情况。在本章中,我们简要解释DNA微阵列的基本原理,包括规划和分析微阵列实验时需要考虑的要点,并以脑膜炎奈瑟菌与人类内皮细胞或上皮细胞的相互作用为例,详细描述其实际应用。