Generalized Schemes for High Throughput Manipulation of the Desulfovibrio vulgaris Hildenborough Genome.
Journal Article
·
· Applied and Environmental Microbiology
more »
- Lawrence Berkeley National Laboratory (LBNL)
- ORNL
- University of Missouri
- University of California, San Francisco
- University of Missouri, Columbia
The ability to conduct advanced functional genomic studies of the thousands of 38 sequenced bacteria has been hampered by the lack of available tools for making high39 throughput chromosomal manipulations in a systematic manner that can be applied across 40 diverse species. In this work, we highlight the use of synthetic biological tools to 41 assemble custom suicide vectors with reusable and interchangeable DNA parts to 42 facilitate chromosomal modification at designated loci. These constructs enable an array 43 of downstream applications including gene replacement and creation of gene fusions with 44 affinity purification or localization tags. We employed this approach to engineer 45 chromosomal modifications in a bacterium that has previously proven difficult to 46 manipulate genetically, Desulfovibrio vulgaris Hildenborough, to generate a library of 47 662 strains. Furthermore, we demonstrate how these modifications can be used for 48 examining metabolic pathways, protein-protein interactions, and protein localization. The 49 ubiquity of suicide constructs in gene replacement throughout biology suggests that this 50 approach can be applied to engineer a broad range of species for a diverse array of 51 systems biological applications and is amenable to high-throughput implementation.
- Research Organization:
- Oak Ridge National Laboratory (ORNL)
- Sponsoring Organization:
- SC USDOE - Office of Science (SC)
- DOE Contract Number:
- AC05-00OR22725
- OSTI ID:
- 1028745
- Journal Information:
- Applied and Environmental Microbiology, Journal Name: Applied and Environmental Microbiology Journal Issue: 21 Vol. 77; ISSN AEMIDF; ISSN 0099-2240
- Country of Publication:
- United States
- Language:
- English
Similar Records
Generalized schemes for high throughput manipulation of the Desulfovibrio vulgaris Hildenborough genome
Toward a rigorous network of protein-protein interactions of the model sulfate reducer Desulfovibrio vulgaris Hildenborough
Journal Article
·
Fri Jul 15 00:00:00 EDT 2011
· Applied&Environmental Microbiology
·
OSTI ID:1052164
Toward a rigorous network of protein-protein interactions of the model sulfate reducer Desulfovibrio vulgaris Hildenborough
Journal Article
·
Sun May 01 00:00:00 EDT 2011
· PLoS One
·
OSTI ID:1051787