High-Level Expression of Truncated Surface Antigen P50 of
            <i>Babesia gibsoni</i>
            in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity

Fukumoto, Shinya; Xu, Xuan; Kadota, Kimie; Igarashi, Ikuo; Sugimoto, Chihiro; Fujisaki, Kozo; Nagasawa, Hideyuki; Mikami, Takeshi; Suzuki, Hiroshi

doi:10.1128/cdli.10.4.596-601.2003

Cited by 15 publications

(21 citation statements)

References 25 publications

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“…Recently, we have succeeded in expressing several protozoan membrane associated antigens, such as SAG1 of N. caninum [4], SAG2 of Toxoplasma gondii [14], EMA-2 of B. equi [13] as soluble proteins in E. coli, by deleting the signal peptide and transmembrane regions. Our previous study shows that the N-terminus aa 1-19 and C-terminus aa 447-466 of P50 functions as signal peptide and transmembrane, respectively [7]. In this study, to obtain high amount of the soluble P50 for practical use in an ELISA, the gene encoding P50t without highly hydrophobic signal peptide and transmembrane regions was expressed in E. coli.…”

Section: Discussionmentioning

confidence: 99%

“…In the previous studies, we have identified and cloned a novel gene encoding a surface antigen with a molecular mass of 50 kDa (P50) by immunoscreening of cDNA library prepared from mRNA of B. gibsoni merozoites with canine infected serum, and demonstrated that P50 is an immunodominant antigen [6,7]. The ELISA with GST-P50f expressed in E. coli was able to diagnose both acute and chronic B. gibsoni infections in dogs [8].…”

Section: Discussionmentioning

confidence: 99%

“…SDS-PAGE and Western blot analyses: SDS-PAGE and Western blot analyses were performed as described previously [7].…”

Section: Parasitementioning

confidence: 99%

“…Recently, Fukumoto et al have identified a novel gene encoding an immunodominat antigen, P50 of B. gibsoni, and evaluated its diagnostic potential in ELISA [6][7][8]. The ELISA with recombinant full-length P50 (P50f) expressed in Escherichia coli was able to differentiate clearly between B. gibsoni-infected do gs and B. canis-infected do gs or noninfected dogs [8].…”

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confidence: 99%

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Serodiagnosis of Babesia gibsoni Infection in Dogs by an Improved Enzyme-Linked Immunosorbent Assay with Recombinant Truncated P50

Verdida

Hara

Xuan

et al. 2004

The Journal of Veterinary Medical Science

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ABSTRACT. The surface antigen P50 of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding truncated P50 (P50t) lacking a signal peptide and C-terminal hydrophobic regions were cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). More than 90% portion of the GST-P50t was expressed as a soluble form, in contrast with GST-P50f (full-len gth), which was completely expressed as an insoluble form. This result indicates that removal of the hydrophobic signal peptide and C-terminus had dramatically improved its hydrophilicity. The purified GST-P50t was tested in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to B. gibsoni in dogs. The ELISA with GST-P50t clearly differentiated between B. gibsoni-infected dog sera and uninfected dog sera. In addition, the ELISA detected no cross-reactivity with sera from dogs experimentally infected with the closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi. Field serum samples collected from dogs in Japan and China were examined for the diagnosis of B. gibsoni infection by using the ELISA. 14.5% (9/62), 5.8% (7/120), and 5.4% (2/37) of tested samples were positive for dogs from Okinawa, Yamaguchi, and Osaka prefectures, Japan, respectively. On the other hand, 4.8% (2/41) of tested samples were positive for dogs from Nanjing, China. These results suggest that the GST-P50t could be a reliabl e reagent for practical use in ELISA for the serodiagnosis of canine piroplasmosis caused by B. gibsoni.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…SDS-PAGE and Western blot analyses: SDS-PAGE and Western blot analyses were performed as described previously [7].…”

Section: Parasitementioning

confidence: 99%

mentioning

confidence: 99%

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Serodiagnosis of Babesia gibsoni Infection in Dogs by an Improved Enzyme-Linked Immunosorbent Assay with Recombinant Truncated P50

Verdida

Hara

Xuan

et al. 2004

The Journal of Veterinary Medical Science

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“…In order to obtain pure and large amounts of P50, we successfully expressed a secretory form of P50 (rP50t) in a culture supernatant of insect cells infected with the recombinant baculovirus by the truncation of the C-terminal anchor region of the transmembrane (6). In a mouse immunization trial, it was confirmed that rP50t retained good immunogenicity (6).…”

mentioning

confidence: 92%

Immunization with Recombinant Surface Antigen P50 of Babesia gibsoni Expressed in Insect Cells Induced Parasite Growth Inhibition in Dogs

Fukumoto

Tamaki

Shirafuji

et al. 2005

Clin Vaccine Immunol

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Molecular characterization and antigenic properties of a novel Babesia gibsoni glutamic acid-rich protein (BgGARP)

Mousa

Cao

Aboge

et al. 2013

Experimental Parasitology

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High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity

Cited by 15 publications

References 25 publications

Serodiagnosis of Babesia gibsoni Infection in Dogs by an Improved Enzyme-Linked Immunosorbent Assay with Recombinant Truncated P50

Serodiagnosis of Babesia gibsoni Infection in Dogs by an Improved Enzyme-Linked Immunosorbent Assay with Recombinant Truncated P50

Immunization with Recombinant Surface Antigen P50 of Babesia gibsoni Expressed in Insect Cells Induced Parasite Growth Inhibition in Dogs

Molecular characterization and antigenic properties of a novel Babesia gibsoni glutamic acid-rich protein (BgGARP)

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