Title page for ETD etd-05042006-164528
|Type of Document
||Cloning and expression of cambialistic Bacteroides fragilis superoxide dismutase gene
||Biochemistry and Nutrition
|Gregory, Eugene M.
|Hess, John L.
|Bevan, David R.
|Johnson, John L.
|Larson, Timothy J.
- Gene expression
- Superoxide dismutase
- Molecular cloning
|Date of Defense
A gene coding for the cambialistic superoxide dismutase
(SOD) was isolated from a LambdaGEM-ll genomic library of
Bacteroides fragilis. In order to generate a complete
genomic library, B. fragilis genomic DNA was partially
digested with the restriction endonuclease Sau3AI and was
ligated to cloning vector, LambdaGEM-ll. After in vitro
packaging, DNA was used to infect E. coli KW 251. The
genomic library was finally established in the plaque
population. Recombinant phage DNAs containing the SOD gene
were detected by a 32P-labelled synthetic oligonucleotide
with 17 bases. The sequence of this oligonucleotide was
deduced from the N-terminal amino acid sequence of B. fragilis FeSOD. Two recombinant phage DNAs were selected
based on he results of plaque hybridization. Further
analysis ith restriction mapping and DNA sequencing
revealed that only one recombinant phage DNA contained the
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