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Volume 147, Issue 1

Cloning and characterization of the gene encoding periplasmic 2′,3′-cyclic phosphodiesterase of O:8 Free

Abstract

The gene encoding periplasmic 2′,3′-cyclic phosphodiesterase in O:8 (designated ), was cloned and expressed in . This enzyme enables to grow on 2′,3′-cAMP as a sole source of carbon and energy. Sequencing and analysis of a 3 kb RI fragment containing the gene revealed an open reading frame of 1179 bp, corresponding to a protein with a molecular mass of 71 kDa. The first 25 amino acid residues show features of a typical prokaryotic signal sequence. The predicted molecular mass of the mature peptide is therefore in agreement with the molecular mass estimated by SDS gel electrophoresis (68 kDa). The putative promoter region contains two possible −10 and −35 regions. Furthermore, the 5′ untranslated region contains sequences with significant homology to the cyclic AMP–cyclic AMP receptor protein binding site and the σ consensus. This region is interrupted by an enterobacterial repetitive intergenic consensus (ERIC) sequence. Deletion of the ERIC element from the promoter region had no effect on expression. In the 3′ untranslated region, a possible rho-independent transcriptional terminator was identified. The deduced amino acid sequence of the CpdB protein shows 76% identity with CpdB of and . CpdB of is exported to the periplasmic space. An isogenic mutant strain, constructed by allelic exchange, was no longer able to grow on 2′,3′-cAMP as sole source of carbon and energy. The CpdB mutant showed no significant change in virulence in an oral and intravenous mouse infection model.

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Keyword(s): 2′,3′-cAMP , cAMP–CRP, cyclic AMP–cyclic AMP receptor protein , cAMP–CRP-binding site , cpdB gene , ERIC , ERIC sequence, enterobacterial repetitive intergenic consensus sequence , NPPC, p-nitrophenyl phosphorylcholine and PNPP, p-nitrophenyl phosphate
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2001-01-01
2024-04-19
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Cloning and characterization of the gene encoding periplasmic 2′,3′-cyclic phosphodiesterase of Yersinia enterocolitica O:8
Microbiology 147, 203 (2001); https://doi.org/10.1099/00221287-147-1-203
/content/journal/micro/10.1099/00221287-147-1-203
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