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Full-Text Articles in Microbiology

Development Of Pyrf-Based Genetic System For Targeted Gene Deletion In Clostridium Thermocellum And Creation Of A Pta Mutant, Shital A. Tripathi, Daniel G. Olson, D. Aaron Argyros, Bethany B. Miller, Trisha F. Barrett, Daniel M. Murphy, Jesse D. Mccool, Anne K. Warner, Vineet B. Rajgarhia, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza Aug 2010

Development Of Pyrf-Based Genetic System For Targeted Gene Deletion In Clostridium Thermocellum And Creation Of A Pta Mutant, Shital A. Tripathi, Daniel G. Olson, D. Aaron Argyros, Bethany B. Miller, Trisha F. Barrett, Daniel M. Murphy, Jesse D. Mccool, Anne K. Warner, Vineet B. Rajgarhia, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza

Dartmouth Scholarship

We report development of a genetic system for making targeted gene knockouts in Clostridium thermocellum, a thermophilic anaerobic bacterium that rapidly solubilizes cellulose. A toxic uracil analog, 5-fluoroorotic acid (5-FOA), was used to select for deletion of the pyrF gene. The ΔpyrF strain is a uracil auxotroph that could be restored to a prototroph via ectopic expression of pyrF from a plasmid, providing a positive genetic selection. Furthermore, 5-FOA was used to select against plasmid-expressed pyrF, creating a negative selection for plasmid loss. This technology was used to delete a gene involved in organic acid production, namely pta, which encodes …


Copper Isotope Fractionation During Surface Adsorption And Intracellular Incorporation By Bacteria, Jesica Urbina Navarrete Jan 2010

Copper Isotope Fractionation During Surface Adsorption And Intracellular Incorporation By Bacteria, Jesica Urbina Navarrete

Open Access Theses & Dissertations

Copper isotopes may prove to be a useful tool for investigating bacteria-metal interactions recorded in natural waters, soils, and rocks. However, experimental data that constrain Cu isotope fractionation in biologic systems are limited and unclear. In this study we utilized Cu isotopes (65Cu) as a tool to investigate Cu-bacteria interactions, including surface adsorption and intracellular incorporation. Experiments were conducted with individual Gram-positive (Bacillus subtilis) and Gram-negative (Escherichia coli) bacterial species as well as with bacterial consortia from several natural environments. Adsorption experiments were conducted with live or dead cells over the pH range 2.5 to 6. Surface adsorption of Cu …