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Diversity of aerobic and facultative alkalitolerant and halotolerant endospore formers in soil from the Alvord Basin,Oregon
Authors:Stephanie A. Smith  James N. Benardini III  Janice L. Strap  Ronald L. Crawford
Affiliation:1. University of Idaho, Environmental Biotechnology Institute, P.O. Box 441052, Moscow, ID 83844-1052, USA;2. University of Ontario Institute of Technology, Ont., Canada L1H 7K4
Abstract:Many proteins produced by Bacillus species isolated from extreme environments have been utilized for industrial purposes, as these extreme environments often promote evolution of unique protein properties. The Borax Lake area is unusual due to its geothermal activity, elevated pH, and high arsenic and salt concentrations in its soils. Soils from this region are likely to harbor alkalitolerant, halotolerant, endospore-forming strains that may be of potential ecological and/or commercial interest. The objectives of this study were to develop new PCR primers that could target Bacillus or closely related 16S rRNA genes, to characterize the diversity of alkalitolerant, halotolerant, endospore-forming organisms in the soils surrounding Borax Lake, and to identify novel organisms that may ultimately provide new enzymes for applied use. A three-pronged approach was used to identify such bacteria in soil samples. Organisms were isolated using two different techniques. Finally, metagenomic DNA from soil samples was subjected to 16S rRNA gene amplification using the newly designed primers. Assays were performed to characterize the halotolerance and alkalitolerance of isolates.
Keywords:HMM, halophilic mineral medium   LB, Luria&ndash  Bertani   PCR-DGGE, PCR-denaturing gel electrophoresis   RFLP, restriction fragment length polymorphism
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