Targeted Gene Disruption by Homologous Recombination in the Oral Spirochete Treponema dentico/a ATCC 35405
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Authors
Hollandsworth, Anne M.
Issue Date
2006
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Treponema denticola has been found to play a significant role in the progression
of periodontal disease based on numerical prevalence, high association level and spatial
location within infected sites. There are a number of virulence factors controlled by
various genes that contribute to the role of T. denticola as a periodontal pathogen. This
investigation attempted to mutate two potentially virulent genes in T. denticola:
TDE1343, a homolog of the htrA/degP gene in E. coli which codes for a heat-shock
protein, and TDE1658, a homolog of the surA gene in E. coli which codes for a
periplasmic peptidyl-prolyl isomerase, using a gene transfer system. The mutation of
TDE1343 involved the disruption of pCF440, which contained an internal fragment of
TDE1343, using a selectable ermF/AM cassette. TDE1658 was mutated by inserting the
ermF/AM cassette into pCF466, which contained a fragment of TOE1658. The results of
this study revealed that the plasmid containing the mutated TDE 1343 was lethal to the
E. coli used in the gene transfer system. However, a plasmid containing the mutated
TDE 1658 was not lethal to E. coli. This study concluded that the method used for gene
transfer in T. denticola is ineffective for constructing a plasmid containing a mutated
TDE1343 gene, but effective for the construction of a plasmid containing a disrupted
TDE1658 gene. However, the incorporation of the mutated TDE1658 into the T.
denticola genome through electroporation and homologous recombination has not yet
been successful.
Description
vii, 41 p.
Citation
Publisher
Kalamazoo College
License
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