Cloning and expression of cambialistic Bacteroides fragilis superoxide dismutase gene
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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 SOD gene.
- Doctoral Dissertations