Korean Journal of Environmental Biology (환경생물)

Korean Society of Environmental Biology (한국환경생물학회)
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Cloning and Overexpression of the Cdd Gene Encoding Cytidine Deaminase from Salmonella typhimurium

  • Published : 2003.03.01
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Abstract

The Salmonella typhimurium cdd gene encoding cytidine deaminase (cyti-dine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5.) was isolated through shotgun clon-ing by complementation of the E. coli odd mutation. By subsequent deletion and sub-cloning from the original 3.7 Kb of EcoRI insert (pSAMI), the precise region of the cdd structural gene is located around the BglII site in the middle part of 1.7 Kb of NruI/PvuI segment. The 1.7 Kb containing odd gene wag subcloned to the pUC18 vector and the nucleotide sequence of the cdd gene was determined. When the putative ribosorne-binding site (Shine-Dalgarno sequence) and initiation codon were predicted to be GAGG at the position 459 and ATG at the position 470, respectively, there was an open reading frame of 885 nucleotides, encoding an 294 amino acid protein. The cdd gene expression in E. coli JF611/pSAMI was amplified about 50 fold compared to that of the wild type. The cdd gene expression was maintained in the stationary phase after rea-ching the peak in the late logarithmic phase.

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References

  1. Nucleic Acids Res. v.7 A rapid alkaline extraction procedure for screening reconbinant plasmid Birnboim HC;J Doly https://doi.org/10.1093/nar/7.6.1513
  2. Kor. J. Appl. Microbiol. Bioeng. v.17 Molecular cloning of Bacillus stearothermophilus cdd gene encoding thermostable cytidind/deoxycytidine deaminase Chang JS;BH Song;JG Kim;SD Hong
  3. Biochem et Biophysica. Acta. v.144 Isolation and characterization of the gene coding for human cytidine deaminase. Demontis S;M Terao;M Brivio;S Zanotta;M Bruschi;E Arattini
  4. Escherichia coli and Salmonella typhimurium;Cellular and Molecular Biology Frederrick FC (et al)
  5. J. Biol. Chem. v.193 Protein meaurement with the folin phenol reagent Lowry OH;NJ Rosebrough;AL Farr;RJ Randall
  6. Nature v.413 Complete sequence of Salmonella enterica serover Typhimurium LT2. McClelland M (et al) https://doi.org/10.1038/35101614
  7. Molecular cloning Maniatis T;EF Fritsch;J Sambrook
  8. Bacterol. Rev. v.34 Pyrimidine metabolism in microorganisms O'donovan GA;J Neuhard
  9. Mol. Gen. Genet. v.216 Chromosomal location, cloning and nucleotide sequence of the Becillus subtilis cdd gene encoding sequence of the Bacillus subtilis cdd gene encoding cytidine/deoxycytidine deaminase Song BH;J Neuhard https://doi.org/10.1007/BF00334391
  10. Protein Expr. Purif. v.15 Cloning, expression, and purification of cytidine deaminase from Arabidopsis thalians Vincenzetti S;C Alessamdra;N Jan;K Schnorr