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Characterization of a leucine aminopeptidase from Toxoplasma gondii OAK
Jia, Honglin; Nishikawa, Yoshifumi; Luo, Yuzi; Yamagishi, Junya; Sugimoto, Chihiro; Xuan, Xuenan.
The M17 family leucine aminopeptidase (LAP) hydrolyze amino acids from the N-terminus of peptides. Many LAPs from parasitic protozoa including Plasmmodium, Trypanosoma and Leishmania, have been intensely investigated because of their crucial roles in parasite biology. In this study, a functional recombinant Toxoplasma gondii LAP (rTgLAP) was expressed in Escherichia coli and its enzymatic activity against synthetic substrates for aminopeptidase, as well as the cellular localization was determined. Our results indicated that TgLAP is a functional aminopeptidase in the cytoplasma of T. gondii.
Palavras-chave: Toxoplasma gondii; Leucine aminopeptidase; Enzymatic activity.
Ano: 2010 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2822
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Expression of truncated Babesia gibsoni thrombospondin-related adhesive proteins in Escherichia coli and evaluation of their diagnostic potential by enzyme-linked immunosorbent assay OAK
Sandagdorj, Narantsatsral; Goo, Youn-Kyoung; Terkawi, Mohamad Alaa; Soma, Takehisa; Luo, Yuzi; Li, Yan; Cao, Shinuo; Aboge, Gabriel Oluga; Nishikawa, Yoshifumi; Badgar, Battsetseg; Xuan, Xuenan.
Among the previously established enzyme-linked immunosorbent assays (ELISAs), an ELISA using the full length of recombinant thrombospondin-related adhesive protein of Babesia gibsoni (rBgTRAPf) is considered as the most sensitive diagnostic method for the detection of antibody to B. gibsoni in dogs. However, the expression of rBgTRAPf in high concentration is poor and thus limits its usefulness as diagnostic antigen. To improve its expression level, we have truncated BgTRAPf into two fragments having either N- or C-terminus (BgTRAPn or BgTRAPc). The expression of BgTRAPc protein in Escherichia coli yielded adequate recombinant protein. The specificity and sensitivity of ELISAs with the truncated proteins were determined using B. gibsoni-experimentally...
Palavras-chave: Babesia gibsoni; Diagnosis; Enzyme-linked immunosorbent assay (ELISA); Thrombospondin-related adhesive protein (TRAP).
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3115
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Molecular and Immunological Characterization of a Novel 32-kDa Secreted Protein of Babesia microti OAK
Ooka, Hideo; Terkawi, Mohamad A; Cao, Shinuo; Aboge, Gabriel; Goo, Youn-Kyoung; Luo, Yuzi; Li, Yan; Nishikawa, Yoshifumi; Igarashi, Ikuo; Xuan, Xuenan; 西川, 義文; 五十嵐, 郁男; 玄, 学南.
A cDNA encoding the Babesia microti 32-kDa protein was identified by serological immunoscreening of a cDNA expression library and designated as BmP32. The full length of BmP32 contains an open reading frame of 918 base pairs consisting of 306 amino acids having a significant homology with B. microti secreted antigen 1. Antiserum raised against recombinant protein (rBmP32) specifically reacted with a 32-kDa native protein of the parasite lysate using western blot analysis. The indirect immunofluorescent antibody test showed a preferable localization of BmP32 in the cytoplasm of the intra- and extracellular parasites. Moreover, BmP32 was secreted in the cytosol of infected erythrocytes, especially during the peak parasitemia and the recovery phase of the...
Ano: 2012 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3820
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Molecular identification and antigenic characterization of a merozoite surface antigen and a secreted antigen of Babesia canis (BcMSA1 and BcSA1) OAK
Zhou, Mo; Cao, Shinuo; Luo, Yuzi; Liu, Mingming; Wang, Guanbo; Moumouni, Paul Franck Adjou; Jirapattharasate, Charoonluk; Iguchi, Aiko; Vudriko, Patrick; Terkawi, Mohamad Alaa; Lowenstein, Mario; Kern, Angela; Nishikawa, Yoshifumi; Suzuki, Hiroshi; Igarashi, Ikuo; Xuan, Xuenan.
Background: Babesia canis is an apicomplexan tick-transmitted hemoprotozoan responsible for causing canine babesiosis in Europe and west Asia. Despite its importance, there is no known rapid diagnostic kit detection of B. canis infection in dogs. The present study identified two novel antigens of B. canis and used the recombinant antigens to establish a rapid, specific and sensitive serodiagnostic technique for detection of B. canis infection. Methods: A complementary DNA (cDNA) expression library was constructed from the mRNA of B. canis and immunoscreened using B. canis-infected dog sera. The cDNAs encoding a merozoite surface antigen and a secreted antigen protein were identified and designated as BcMSA1 and BcSA1, respectively. The recombinant BcMSA1...
Palavras-chave: Babesia canis; Canine babesiosis; BcMSA1; BcSA1; ELISA; Immunochromatographic tests.
Ano: 2016 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4420
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