Title page for ETD etd-11072008-063340
|Type of Document
||Lampe, Robert Carl
||The 16S rRNA characterization of a novel "microaerophilic" Pseudomonas sp. from the oligotrophic deep subsurface environment
||Master of Science
|Benoit, Robert E.
|Krieg, Noel R.
|Yousten, Allan A.
- DNA reassocation
- 16S rRNA
|Date of Defense
A gram negative microaerophilic bacterium, designated Pseudomonas sp. strain
MR 100, was isolated from a depth of 463 meters at the Savannah River DOE site and
identified using 16S rDNA sequencing and DNA-DNA reassociation. Micro aerophiles
from the Middendorf formation were isolated by use of a semi-solid agar assay, and
constituted 10% of the plateable microorganisms. Genetic identification involved the
isolation of genomic DNA and amplification of the gene encoding 16S rRNA by PCR,
using universal primers. The amplified DNA was sequenced and compared to 16S rRNA
sequences in Genbank. High sequence similarity (98.5%) was observed with the
Pseudomonas mendocina type strain, indicating a similarity to the (Group I)
pseudomonads. DNA-DNA reassociation was performed between Pseudomonas sp.
strain MR 100 and 11 representative p seudomonads using the S 1 nuclease method. Strain
MR 100 was found to be 20% homologous to the Pseudomonas mendocina type strain,
10% homologous to Pseudomonas alcaligenes, and 5% homologous to Pseudomonas
aeruginosa. Data from biochemical tests confirm the hypothesis that strain MR 100 is a
novel species of Pseudomonas
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