Lab-based ENIGMA researchers enrich, isolate and physiologically/genetically characterize the specific taxa that are predicted to be the most important to the observed field processes. By characterizing genetic functions and linking genotype to phenotype in the laboratory, we can interpret field data mechanistically, leading to pointed hypotheses of the role of particular taxa, proteins, co-factors, and their interactions in creating systems behavior.
Relevant Publications
Genome Editing in Rhodanobacter denitrificans - ENIGMA researchers demonstrated the development and application of a markerless deletion mutagenesis system in nitrate-reducing bacterium Rhodanobacter denitrificans. This method marks a crucial step in advancing Rhodanobacter as a model denitrifying bacterium for the study of denitrification in groundwater ecosystems and diverse molecular mechanisms of low-pH resistance. More →
Two model phages characterized by new CRISPR based technology extensible to diverse phages - ENIGMA researchers demonstrated for the first time that they can, on a genome-wide scale, identify phage genes that are essential (or not) to infecting bacteria, and then replace non-essential DNA with distinctive barcode tags. Their method could unlock potent biotechnology applications. More →
Novel Protocol Leverages Automatic Liquid Transfer to Prepare Hundreds of Microbial Cell Cultures for Proteomic Analysis - ENIGMA researchers detailed a step-by-step protocol that consists of cell lysis in alkaline chemical buffer (NaOH/SDS) followed by protein precipitation with high-ionic strength acetone in 96-well format. This protocol, combined with previously established automated protein quantification and protein normalization protocols, provides a rapid, cost-effective method to prepare LC-MS proteomic samples from bacteria and non-filamentous fungi cell cultures. More →
Microbial Communities Vary Widely Underground, Even When Close - Study finds that geochemistry and hydrogeology can influence community type and function, even when those communities are only 10 cm apart. More →
Multicopy suppressor screens reveal convergent evolution of single-gene lysis proteins - Using new barcoded screening technology, ENIGMA researchers uncovered genome-wide targets of diverse including single-stranded RNA-phages (ssRNA-phages)-derived protein antibiotics to efficiently identify the host mechanisms targeted in bacteriolysis. The novel approach could have significant applications for characterizing the hundreds of putative Sgl identified in genomic databases and recent studies. More →
Examining adaptations of a dominant bacterial species at a contaminated subsurface site - Leveraging comparative genomics, community analysis, and geochemical data to understand adaptations that facilitate survival of a dominant community member in the contaminated Oak Ridge Reservation subsurface. More →