Unlike other bacteria, C. difficile must detect both a germinant signal and a co-germinant signal to trigger spore germination, but how these cues are sensed is unknown. This study finds that the CspC...

Linezolid-resistant Clostridioides difficile, conferred by the acquisition of cfr-like genes, has been reported in Europe and America. However, the emergence of linezolid resistance in C. difficile...

Intranasal CR9114 is safe in humans, and twice-daily dosing confers protection against influenza virus challenge in macaques.

The gut microbiota communicates with multiple organs through neuroactive molecules, but the specific compounds produced remain unclear. We investigate…

This is the twelfth minor release (v1.12) providing more than 10 minor improvements and many bug fixes improving runtime stability, IO compatibility, and last but not least user experience. Compati...

In this Review, Castañeda-Barba, Top and Stalder use the One Health framework to synthesize the recent literature on the ecological and evolutionary factors that determine the successful local and glo...

Abstract. Bidirectional replication is a conserved principle requiring coordinated translocation of the two replicative helicases loaded at the origin. In

Biologics are delivered to the gut using phage that infects resident commensal bacteria.

Protein structure is conserved beyond sequence, making multiple structural alignment (MSTA) essential for analyzing distantly related proteins. Computational prediction methods have vastly extended ou...

This study finds that native tree extinctions and alien naturalizations are pushing forests towards fast-growing, resource-demanding species. This global shift could affect carbon storage and ecosyste...

Here, the authors show that packaging of bacterial DNA by phage-like particles is widespread in the gut microbiome, with activity of gene transfer agents being prominent in Oscillospiraceae and Rumino...

We used whole-genome sequencing to study Clostridioides difficile isolates from patients with recurrent or sequential infections in Minnesota, USA. We summ

Combinatorial DNA design and assembly is an efficient and pragmatic way to obtain high-performing metabolic pathway designs quickly. However, implementation may require organism-specific technical barriers to be overcome. Firstly, suitable expression control parts such as promoters and ribosome-binding sites (RBSs) which provide a suitable range of expression levels need to be identified or developed. Secondly, these need to be assembled into pathway-encoding combinatorial libraries of sufficient size, quality and diversity. For organisms with transformation frequencies too low to allow direct transformation of library assembly reactions, such as many Clostridium spp., assembly and amplification is typically carried out using Escherichia coli. However, if constructs are deleterious (or 'burdensome') to E. coli, which is often the case when using Clostridium genetic parts, poor libraries may be obtained. Here we develop a new approach called integration‑coupled activation of promoterless sequences (ICAPS) to overcome this barrier and therefore enable combinatorial assembly in Clostridium. Libraries were designed and assembled as promoterless synthetic operons, preventing expression during DNA assembly, and expression was only activated later, when constructs were integrated into the host genome downstream of a promoter. Variation of expression levels was achieved using a range of context-resistant RBS sequences. This approach was used to produce a Clostridium acetobutylicum library with combinatorial expression variants of an introduced hexanol pathway. This proof of concept provides a generally-applicable approach to implement combinatorial metabolic pathway-encoding libraries in Clostridium spp., circumventing the excessive strength of Clostridium expression control parts in E. coli, and is applicable to other organisms. ### Competing Interest Statement The authors have declared no competing interest. BBSRC, BB/M002454/1, BB/V001396/1