Project description:Ammonia-oxidizing archaeal (AOA) amoA diversity and relative abundance in Gulf of Mexico sediments (0-2 cm) were investigated using a functional gene microarray; a two color array with a universal internal standard
Project description:Expression data for Desulfovibrio alaskensis strain G20 and mutants in regulator proteins grown on lactate sulfate media and then pelleted and transferred to another media when they reached stationary phase. The Choline mutant was transferred to lacte/sulfate minimal media and choline/sulfate minimal media. The LysX mutant was transferred to minimal media with lysine and rich media. G20 was transferred to minimal media, choline/sulfate minimal media, lactate/choline/sulfate minimal media, minimal media with lysine, and rich media. We aimed to confirm or expand the regulons of each of the transposon interupted regulator mutants and compare gene expression responses of the regulators in different growth conditions. 10 samples were collected: 2 regulator mutants (2 conditions each), Desulfovibrio alaskensis G20 (5 conditions), 2 replicates for G20 minimal media condition. Control sample -G20 rich media.
Project description:Expression data for Desulfovibrio alaskensis strain G20 grown on lactate in sulfate-limited monoculture and syntrophic coculture with Methanococcus maripaludis or Methanospirillum hungatei in chemostats at a low growth rate of 0.027h-1. 7 samples of Desulfovibrio alaskensis strain G20 grown in syntrophic coculture on lactate with either Methanococcus maripaludis (4 replicates) or Methanospirillum hungatei (3 replicates), and 5 samples of sulfate-limited monoculture growth of strain G20 on lactate.
Project description:Ammonia-oxidizing archaeal (AOA) amoA diversity and relative abundance in Gulf of Mexico sediments (0-2 cm) were investigated using a functional gene microarray; a two color array with a universal internal standard Two color array (cy3 and cy5): the universal standard 20 bp oligo (fluoresced with cy5) is printed to the slide with a 70-mer. Environmental DNA sequences (fluoresced with Cy3) within 15% of the 70-mer will bind to it. Signal is the cy3/cy5. Up to four arrays per sample, with two biological replicates made into two targets, each run on duplicate arrays.
Project description:In order to genetically determine the essential components of electron flow during respiration of sulfate by the anaerobe Desulfovibrio desulfuricans G20, a growth mode independent of sulfate is needed. However, a mutant of the G20 strain, I2, lacking the abundant type-1 tetraheme cytochrome c3 (TpI-c3), was found to be altered in growth with sulfate. Here we report that I2 does not reduce sulfate with electrons from pyruvate. In contrast, in the absence of sulfate, fermentative growth of I2 on pyruvate actually yielded slightly greater cellular protein than G20. When provided lactate with sulfate or when fermenting pyruvate, I2 produced more hydrogen and accumulated little if any succinate. G20 grows by fumarate dismutation and accumulates the theoretical 2:1 ratio of the end products, succinate and acetate, respectively. In contrast, I2 did not grow on fumarate alone. We inferred that TpI-c3 might be required for transfer of electrons to the membrane-bound fumarate reductase or for establishing a redox environment needed to trigger synthesis of the enzymes for succinate production. Microarray and proteomic analyses of gene and protein expression in I2 compared with G20 were consistent with increases in those enzymes predicted to be necessary for the generation of end products measured. These results support a role for the periplasmic TpI-c3 in electron flow from pyruvate to sulfate and in establishing conditions for enzyme production for fumarate dismutation. In addition, evidence for the simultaneous operation of respiratory electron flow and substrate-level phosphorylation was obtained from end product analysis. Strains I2 and G20 of Desulfovibrio desulfuricans were grown in different media. Four biological replicates of G20 on lactate media, and two biological replicates of G20 on Pyruvate media were compared with three replicates of I2 on lactate media and two replicates of I2 on pyruvate media.
