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Regulation of Corynebacterium ammoniagenes purF and Isolation of purF-Specific Regulatory Proteins  

Lee, Seok-Myung (Department of Biotechnology and Bioinformatics, Korea University)
Kim, Youn-Hee (Department of Oriental Medicine, Semyung University)
Lee, Heung-Shick (Department of Biotechnology and Bioinformatics, Korea University)
Publication Information
Korean Journal of Microbiology / v.45, no.3, 2009 , pp. 233-238 More about this Journal
Abstract
The expression of Corynebacterium ammoniagenes purF was analyzed by utilizing a plasmid carrying a cat gene fused to the purF promoter region. Adenine and guanine repressed the expression of the purF gene by 20~30% but hypoxanthine did not exert such repressive effect. The expression purF was maximal at the late log phase and remained constant throughout the stationary phase. Promoter $P_{180}$ which was developed in C. glutamicum was also functional in C. ammoniagenes, achieving maximal activity at the late log phase. The promoter outperformed Escherichia coli $P_{tac}$ promoter by 40~50% level. DNA-affinity purification identified a protein which could bind to the promoter region of the purF gene. The protein showed high similarity to the CRP-family transcriptional regulator encoded by NCgl0120 in C. glutamicum. The size of the screened protein agreed with the expected protein size from the ORF NCgl0120. The corresponding gene in C. ammoniagenes encoded a 42 kDa polypeptide composed of 400 amino acids with expected pI of 4.9. The encoded protein showed 14.1% and 15.8% identity with E. coli and Bacillus subtilis PurR, respectively, suggesting that the isolated protein might be a novel type of regulatory protein involved in the regulation of purine metabolism.
Keywords
C. ammoniagenes; purF; purine biosynthesis;
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