Department of Microbiology and Alimentary Pharmabiotic Centre, National University of Ireland, Cork, Ireland.
PLoS One. 2013 May 30;8(5):e64699. doi: 10.1371/journal.pone.0064699. Print 2013.
Bifidobacteria are claimed to contribute positively to human health through a range of beneficial or probiotic activities, including amelioration of gastrointestinal and metabolic disorders, and therefore this particular group of gastrointestinal commensals has enjoyed increasing industrial and scientific attention in recent years. However, the molecular mechanisms underlying these probiotic mechanisms are still largely unknown, mainly due to the fact that molecular tools for bifidobacteria are rather poorly developed, with many strains lacking genetic accessibility. In this work, we describe the generation of transposon insertion mutants in two bifidobacterial strains, B. breve UCC2003 and B. breve NCFB2258. We also report the creation of the first transposon mutant library in a bifidobacterial strain, employing B. breve UCC2003 and a Tn5-based transposome strategy. The library was found to be composed of clones containing single transposon insertions which appear to be randomly distributed along the genome. The usefulness of the library to perform phenotypic screenings was confirmed through identification and analysis of mutants defective in D-galactose, D-lactose or pullulan utilization abilities.
双歧杆菌被认为通过一系列有益的或益生菌的活性对人体健康有积极的影响，包括改善胃肠道和代谢紊乱，因此近年来，这组特殊的胃肠道共生菌受到了越来越多的工业和科学关注。然而，这些益生菌机制的分子机制在很大程度上仍然未知，主要是因为双歧杆菌的分子工具发展得相当不完善，许多菌株缺乏遗传可及性。在这项工作中，我们描述了两种双歧杆菌菌株，即短双歧杆菌 UCC2003 和短双歧杆菌 NCFB2258 的转座子插入突变体的产生。我们还报告了双歧杆菌菌株中转座子突变体文库的创建，该文库使用了短双歧杆菌 UCC2003 和基于 Tn5 的转座酶策略。该文库由含有单个转座子插入的克隆组成，这些插入似乎随机分布在基因组中。该文库通过鉴定和分析在半乳糖、D-乳糖或普鲁兰利用能力上有缺陷的突变体，证实了其用于表型筛选的有用性。
