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KMID : 0545120000100050700
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Journal of Microbiology and Biotechnology
2000 Volume.10 No. 5 p.700 ~ p.706
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Cloning, DNA Sequence Determination, and Analysis of Growth-Associated Expression of sodF Gene Coding for Fe-and Zn-Containing Superoxide Dismutase of Streptomyces griseus
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Kim Ju-Sim
Lee Jeong-Kug
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Abstract
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Iron- and zinc-containing superoxide dismutase (FeZnSOD) and nickel-containing superoxide dismutase (NiSOD) are cytoplasmic enzymes in Streptomyces griseus. The sodF gene coding for FeZnSOD was cloned from genomic Southern hybridization analysis with a 0.5-kb DNA probe, which was PCR-amplified with facing primers corresponding to the N-terminal amino acids of the purified FeZnSOD of S. griseus and a C-terminal region which is conserved among bacterial FeSODs and MnSODs. The sodF open reading frame (ORF) was comprised of 213 amino acids (22,430Da), and the deduced sequence of the protein was highly homologous (86% identity) to that of FeZnSOD of Streptomyces coelicolor. The FeZnSOD expression of exponentially growing S. griseus cell was approximately doubled as the cell growth reached the early stationary phase. The growth-associated expression of FeZnSOD was mainly controlled at the transcriptional level, and the regulation was exerted through the 110bp regulatory DNA upstream from the ATG initiation codon of the sodF gene.
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