细菌的功能基因组学通常旨在建立工业微生物中的基因型-表型联系，技术和生物医学的相关性。在这方面,随机转座子诱变与高通量下一代测序方法相结合，称为转座子插入测序（TIS），已经成为一个强大的，全基因组替代进行功能基因组分析。尽管这些方法已用于涉及致病性和临床相关细菌的大量研究中，它们在共生细菌和潜在有益细菌领域很少受到关注，包括益生菌微生物。在这一章中,我们描述了TIS方法转座子定向插入测序的实施,以描述包含几种共生和潜在益生菌的属的代表性菌株中的必需基因集,并讨论了将类似的方法学方法应用于其他感兴趣的双歧杆菌物种/菌株时的注意事项。
Functional genomics of bacteria commonly aims at establishing genotype-phenotype links in microorganisms of industrial, technological and biomedical relevance. In this regard, random transposon mutagenesis coupled to high-throughput next-generation sequencing approaches, termed transposon-insertion sequencing (TIS), has emerged as a robust, genome-wide alternative to perform functional genome analysis. Though these approaches have been used in a large number of studies involving pathogenic and clinically relevant bacteria, they have received little attention in the fields of commensal and potentially beneficial bacteria, including probiotic microorganisms. In this chapter, we describe the implementation of the TIS method Transposon-Directed Insertion Sequencing to describe the set of essential genes in a representative strain of a genus encompassing several commensal and potentially probiotic bacteria and discuss considerations when applying similar methodological approaches to other Bifidobacterium species/strains of interest.
