Marine Biotechnology Fellowship: Molecular Genetics of Ammonium Assimilation in the Oligotrophic Marine Bacterium, RM-1

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9321698 CIFUENTES This Research Fellowship in Marine Biotechnology will investigate the molecular genetics and biochemistry of the enzyme, glutamine synthetase in a true marine bacterium to better understand nitrogen metabolism of marine bacterioplankton. The glnA gene will be cloned from the marine bacterium strain RM-1 and sequenced. RM-1, an aerobic obligate oligrotrophic bacterium, was isolated using dilution culture techniques by D Button of the University of Alaska, Fairbanks. This technique helps isolate the most abundant strain rather than those which are most easily cultured in the laboratory. If the cloning and sequencing of glnA proceeds on schedule, than RM-1 glutamine synthetase expressed in an E. coli mutant will be purified and used to generate anitsera for probing of the glutamine synthetase activity in natural samples. Probes of metabolic function developed in this Fellowship will be applied to an on-going collaborative study of nitrogen biogeochemistry in the Gulf of Mexico. Understanding the molecular genetics of a key enzyme in the nitrogen metabolism of a typical marine bacterium will provide new information on physiological function in oligotrophic environments and improve the understanding of important ecological interactions and biogeochemical processes in N-cycling. ***