Cloning and Expression of Bartonella henselae sucB Gene Encoding an Immunogenic Dihydrolipoamide Succinyltransferase Homologous Protein

Abstract

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 BamHI 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 (sucB). 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.

Identification of immunogenic proteins of Flavobacterium columnare by two‐dimensional electrophoresis immunoblotting with antibacterial sera from grass carp, Ctenopharyngodon idella (Valenciennes)

“…Several other proteins identified as immunogens in this study have also been reported as such in other pathogenic microorganisms, such as dihydrolipoamide succinyltransferase in Coxiella burnetti (Nguyen, To, Yamaguchi, Fukushi & Hirai 1999) and Bartonella henselae (Kabeya, Maruyama, Hirano & Mikami 2003), alcohol dehydrogenase in Candida albicans (Bertram, Swoboda, Gooday, Gow & Brown 1996), beta subunit of succinyl-CoA synthetase in Leptospira interrogans and L. borgpetersenii (Sakolvaree, Maneewatch, Jiemsup, Klaysing, Tongtawe, Srimanote, Saengjaruk & Chaicumpa 2007), elongation factor G in Lactococcus garvieae (Shin, Palaksha, Kim, Nho, Cho, Heo, Heo, Park & Jung 2007) and fructosebisphosphate aldolase, which can elicit protective immunity against group A streptococcus challenge in mice (Ma, Li, Wang, Zeng, Yin, Feng & Wei 2009).…”

Proteomic and Immunoblot Analyses of Bartonella quintana Total Membrane Proteins Identify Antigens Recognized by Sera from Infected Patients

“…SucB (dihydrolipoamide succinyltransferase), a 43.8-kDa cytosolic protein, was recognized by 76% of our patient sera (Table 4) and has been detected in immunoscreens of genomic expression libraries for both B. henselae and Bartonella vinsonii subsp. berkhoffii (15,18,26). Another cytosolic protein, the 63.8-kDa protein FtsZ (spots JB48 and JB56), reacted with sera from 24% of our patients and has been identified previously as a potential diagnostic antigen for Bartonella infection (19,20,36).…”

Bartonella henselae Recombinant Pap31 for the Diagnosis of Canine and Human Bartonelloses

“…Currently, the diagnosis of canine and human Bartonelloses is performed by the isolation of Bartonella by culture, the amplification of Bartonella DNA by PCR, and the detection of Bartonella antibodies by serological assays. Although serology can only confirm exposure, immunofluorescent antibody assays (IFAs), Western Blot (WB), and enzyme-linked immunosorbent assays (ELISAs) are most frequently used for the diagnosis of canine and human Bartonelloses [11][12][13][14][15][16][17][18][19][20]. Previously, we reported that the sensitivity of IFA did not substantially improve despite using a panel consisting of eight Bartonella IFA antigens, each tested as an independent serological assay [21].…”