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Cloning and Expression of Bartonella henselae suc B Gene Encoding an Immunogenic Dihydrolipoamide Succinyltransferase Homologous Protein
Author(s) -
Kabeya Hidenori,
Maruyama Soichi,
Hirano Kouji,
Mikami Takeshi
Publication year - 2003
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2003.tb03419.x
Subject(s) - immunoscreening , biology , microbiology and biotechnology , bartonella henselae , brucella melitensis , genomic dna , recombinant dna , gene , sequence analysis , genomic library , nucleic acid sequence , virology , fusion protein , xhoi , bamhi , peptide sequence , genetics , brucella , serology , cdna library , antibody , brucellosis
Immunoscreening of a ZAP genomic library of Bartonella henselae strain Houston‐1 expressed in Escherichia coli resulted in the isolation of a clone containing 3.5 kb Bam HI genomic DNA fragment. This 3.5 kb DNA fragment was found to contain a sequence of a gene encoding a protein with significant homology to the dihydrolipoamide succinyltransferase of Brucella melitensis ( suc B). Subsequent cloning and DNA sequence analysis revealed that the deduced amino acid sequence from the cloned gene showed 66.5% identity to SucB protein of B. melitensis , and 43.4 and 47.2% identities to those of Coxiella burnetii and E. coli , respectively. The gene was expressed as a His‐Nus A‐tagged fusion protein. The recombinant SucB protein (rSucB) was shown to be an immunoreactive protein of about 115 kDa by Western blot analysis with sera from B. henselae ‐immunized mice. Therefore the rSucB may be a candidate antigen for a specific serological diagnosis of B. henselae infection.