This section includes protocols we have developed and published (in more detail than can be provided in journal articles), as well as computational resources and a list of plasmids and strains we have developed and are happy to share with the broader community. We only ask that if these resources are useful to you, please cite our work.
The extracellular polymeric substance produced by E. limosum can make this organism very difficult to work with. We have generated three different knockout strains:
We generated a range of E. limosum vectors for constitutive or inducible expression (Sanford and Woolston, 2022). Promoters include:
We also developed new selectable markers, including:
As part of a project supported by the DOE Joint Genome Institute's Community Science Program, we synthesized a library of 57 RecT homologs for screening in Clostridia, codon optimized for expression in Eubacterium limosum. These are all on plasmids with the pIP404 ori, chloramphenicol resistance gene (catP), and with expression controlled by the p2TetO1 promoter originally developed for Clostridium acetobutylicum. As such, they should be broadly compatible with different Clostridial species. The whole library or individual plasmids are available upon request
Reference: Sanford et al, 2024, In Review
Recombinase Sequences (xlsx)
DownloadTo request any of the above strains (or other published material not listed above), contact Prof. Woolston using the link below. Please include your shipping address, as well as a FedEx Account to cover shipping costs. If shipping costs are prohibitive, please let us know, and we will do our best to accommodate.
All computer code used to support analysis and conclusions in our published work is available on our GitHub repository
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