Cloning and characterization of putative zinc protease genes of Ehrlichia canis

Ching-Hao Teng, Stephen C. Barr, Yung Fu Chang

Research output: Contribution to journalArticle

Abstract

A putative zinc protease gene operon from Ehrlichia canis was cloned and sequenced. A genomic library was constructed in a pHG165 plasmid vector using Sau3A partially digested E. canis chromosomal DNA. Sequence analysis of the insert DNA from this clone indicated two open reading frames with a size of 1314 and 1350bp that encodes for ProA and ProB, respectively. Based on BLAST analyses, ProA and ProB share 20-30% identities with members of the eukaryotic mitochondrial processing peptidase (MPP) subfamily, which are heterodimers containing α and β subunits. The subunits share a 20% of identity, but only MPP-β contains a conserved zincbinding motif, His-Xaa-Xaa-Glu-His (HXXEH). proA and proB are also detectable in E. canis and Ehrlichia chaffeensis, but not the Anaplasma phagocytophila. 5′-RACE revealed that the 5′ end of the proA mRNA is heterogeneous, containing additional adenine residues that may be directed by pseudo-templated transcription. Although ProA was identified in both E. canis and E. chaffeensis, ProB was detected only in E. canis. ProA and ProB were both detectable in E. canis-infected DH82 cells. Sera from dogs, which were either naturally or experimentally infected with E. canis, recognized both the recombinant protein antigens.

Original languageEnglish
Pages (from-to)109-121
Number of pages13
JournalDNA Sequence - Journal of DNA Sequencing and Mapping
Volume14
Issue number2
DOIs
Publication statusPublished - 2003 Apr 1

Fingerprint

Ehrlichia canis
Cloning
Zinc
Organism Cloning
Peptide Hydrolases
Genes
Genomic Library
DNA
Adenine
Transcription
Recombinant Proteins
Ehrlichia chaffeensis
Plasmids
Antigens
Messenger RNA
Anaplasma phagocytophilum
Operon
mitochondrial processing peptidase
DNA Sequence Analysis
Open Reading Frames

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Genetics
  • Endocrinology

Cite this

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abstract = "A putative zinc protease gene operon from Ehrlichia canis was cloned and sequenced. A genomic library was constructed in a pHG165 plasmid vector using Sau3A partially digested E. canis chromosomal DNA. Sequence analysis of the insert DNA from this clone indicated two open reading frames with a size of 1314 and 1350bp that encodes for ProA and ProB, respectively. Based on BLAST analyses, ProA and ProB share 20-30{\%} identities with members of the eukaryotic mitochondrial processing peptidase (MPP) subfamily, which are heterodimers containing α and β subunits. The subunits share a 20{\%} of identity, but only MPP-β contains a conserved zincbinding motif, His-Xaa-Xaa-Glu-His (HXXEH). proA and proB are also detectable in E. canis and Ehrlichia chaffeensis, but not the Anaplasma phagocytophila. 5′-RACE revealed that the 5′ end of the proA mRNA is heterogeneous, containing additional adenine residues that may be directed by pseudo-templated transcription. Although ProA was identified in both E. canis and E. chaffeensis, ProB was detected only in E. canis. ProA and ProB were both detectable in E. canis-infected DH82 cells. Sera from dogs, which were either naturally or experimentally infected with E. canis, recognized both the recombinant protein antigens.",
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Cloning and characterization of putative zinc protease genes of Ehrlichia canis. / Teng, Ching-Hao; Barr, Stephen C.; Chang, Yung Fu.

In: DNA Sequence - Journal of DNA Sequencing and Mapping, Vol. 14, No. 2, 01.04.2003, p. 109-121.

Research output: Contribution to journalArticle

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