TY - JOUR
T1 - Cloning and characterization of putative zinc protease genes of Ehrlichia canis
AU - Teng, Ching Hao
AU - Barr, Stephen C.
AU - Chang, Yung Fu
N1 - Funding Information:
This work was supported by grants from the Fort Dodge Animal Health, a Division of American Home Products Corporation and the New York State Science and Technology Foundation. We thank to Dr Lidstrom for her gifts of Methylobacterium extorquens AM1 pqqE and pqqF mutants, the mobilizing plasmid pRK2013, and pRK310.
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2003/4
Y1 - 2003/4
N2 - 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.
AB - 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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U2 - 10.1080/1042517031000073736
DO - 10.1080/1042517031000073736
M3 - Article
C2 - 12825352
AN - SCOPUS:0037382852
VL - 14
SP - 109
EP - 121
JO - DNA Sequence - Journal of DNA Sequencing and Mapping
JF - DNA Sequence - Journal of DNA Sequencing and Mapping
SN - 1042-5179
IS - 2
ER -