This study is interested in isolating and characterizing bacteriophages from local water environments. The methods of isolating these bacterial strain-specific viruses need to be further developed. Suggested action to improve viral isolation includes isolating phages using bacteria other than E.coli and determining the medium that best supports phage growth. Also, this study is interested in understanding what conditions best support the swarming phenotype typical to bacterial cells present in biofilm formations. The first step in this process is developing a method for observing the swarming form of motility. Additionally, it is the goal of this research to further characterize the swarming phenotype by isolating and examining the genomic DNA of a transposon mutant that exhibits the positive swarming behavior.
The results of this study indicate that the method most successful in isolating bacteriophages from local water environments involves the use of 1X soft-top agar supplemented with MgSO4 and CaSO4 (1% (w/v) proteose peptone, 0.5% (w/v) yeast extract, 43mM NaCl, 46mM K2HPO4, 0.7% (w/v) agar, 0.5mM MgSO4, 0.5mM CaSO4). From this improvement, viruses were more consistently isolated, but further research is still necessary to characterize and identify these viral strains.
The results of the swarming project show that succinate as a carbon source does not support the swarming phenotype, nor does maleic acid. Glutamate as the sole carbon source in the environment supported swarming activity in all strains, but in the presence of other carbon source, the swarming activity of each strain varied greatly. Further research is necessary to better determine how each strain behaves in each environment.
The results of the DNA isolation of ME4, a mutant containing the Tn5 transposon, indicate that the developed polymerase chain reaction procedure is acceptable for isolating an amplified fragment of approximately 200 base pairs. Further research is necessary to sequence this fragment and determine if the swarming phenotype is contained within this region.
Britton Walker, ’07 Ft. Collins, CO
Majors: Biochemistry and Molecular Biology, Spanish, Chemistry
Sponsor: Jeffrey Cardon