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Chromosomal location, cloning and nucleotide sequence of the Bacillus subtilis cdd gene encoding cytidine/deoxycytidine deaminase

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Summary

The Bacillus subtilis cdd gene encoding cytidine/2′-deoxycytidine deaminase has been located by transduction at approximately 225 degrees on the chromosome, and the gene order rpC-lys-cdd-aroD was established. The gene was isolated from a library of B. subtilis DNA cloned in λD69 by complementation of an Escherichia coli cdd mutation. Minicell experiments revealed a molecular mass of 14000 dalton for the cytidine deaminase subunit encoded by the cloned DNA fragment. The molecular weight of the native enzyme was determined to be 58000, suggesting that it consists of four identical subunits. The nucleotide sequence of 1170 bp, including the cdd gene, was determined. An open reading frame encoding a polypeptide with a calculated molecular mass of 14800 dalton was deduced to be the coding region for cdd. The deduced amino acid composition of the 136-amino acid-long subunit shows that it contains six cysteine residues. A computer search in the GenBank DNA sequence library revealed that the 476 bp HindIII fragment containing the putative promoter region and the first ten codons of cdd is identical to the P43 promoter-containing fragment previously isolated by Wang and Doi (1984). They showed that the fragment contained overlapping promoters transcribed by B. subtilis δ43 and δ37 RNA polymerase holoenzymes during growth and stationary phase.

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Abbreviations

SDS:

sodium dodecyl sulphate

Ap:

ampicillin resistance

Tetr :

tetracycline resistance

Kmr :

kanamycin resistance

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Author notes

  1. Bang-Ho Song

    Present address: Department of Biology, Teachers College, Kyung-pook National University, 702-701, Taegu, South Korea

Authors and Affiliations

  1. Enzyme Division, Institute of Biological Chemistry B, University of Copenhagen, Sølvgade 83, DK-1307, Copenhagen K, Denmark

    Bang-Ho Song & Jan Neuhard

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  1. Bang-Ho Song
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  2. Jan Neuhard
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Communicated by H. Hennecke

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Song, BH., Neuhard, J. Chromosomal location, cloning and nucleotide sequence of the Bacillus subtilis cdd gene encoding cytidine/deoxycytidine deaminase. Mol Gen Genet 216, 462–468 (1989). https://doi.org/10.1007/BF00334391

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  • DOI: https://doi.org/10.1007/BF00334391

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