Research Highlights for Milestone 1: Sequences, Proteins, Molecular Complexes
- Microbial genome sequencing at DOE has produced the sequences of over 200 microbes (see Microbial Genomes Sequenced or in Process by DOE).
- Microbial communities are being sequenced from environments as diverse as acid mine drainage sites (where the pH is less than 1.0) and the Sargasso Sea (e.g., Metagenomics: Opening a New Window onto Natural Microbial Communities).
- Single-cell methods are being developed to sequence individual organisms from complex communities, and a number of laboratory culture-independent approaches are being used to investigate the composition and functionality of microbial communities.
- Improved methods for synthesizing genomes are being developed to test our understanding of gene function and regulation (see Accurate, Low-Cost Gene Synthesis from Programmable DNA Microchips).
- Several projects are developing new concepts and strategies to generate recalcitrant proteins such as those in membranes and those containing metals.
- Others are piloting high-throughput methods for turning out proteins needed now by GTL projects. The ultimate goal is creating the capabilities to produce on demand any protein potentially expressed by microbes and microbial communities (see Facility for Production and Characterization of Proteins and Molecular Tags).
- Some GTL projects are developing methods for creating new classes of affinity reagents for high-throughput global assays (e.g., on chips) of protein-expression and interaction partners to identify, track, remove, and disable corresponding proteins; locate protein complexes in living systems; and for other purposes (see Molecular Tags: Fusion Tags and Affinity Reagents).
- Technologies are being refined, validated, and deployed in increasingly automated pilot pipelines to cultivate, isolate, stabilize, and characterize molecular complexes by making use of miniaturization and other developments, focusing on a number of microbes including Rhodopseudomonas and Shewanella (e.g., Facility for Production and Characterization of Proteins and Molecular Tags and Capturing and Characterizing Protein Complexes, the Workhorses of the Cell).
- Live-cell imaging, including colocalization and FRET-based techniques, are being used to observe complexes.
- Capabilities for modeling molecular-machine shapes and reaction surfaces are being developed and tested.
