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Establishment of a two‐dimensional electrophoresis map for Neospora caninum tachyzoites by proteomics
Author(s) -
Lee Eung Goo,
Kim Jae Hoon,
Shin Yong Seung,
Shin Gee Wook,
Suh Myung Deuk,
Kim Dae Yong,
Kim Yong Hwan,
Kim Gon Sup,
Jung Tae Sung
Publication year - 2003
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300617
Subject(s) - neospora caninum , triosephosphate isomerase , biology , microbiology and biotechnology , two dimensional gel electrophoresis , antigen , proteomics , proteome , peptide mass fingerprinting , gel electrophoresis , biochemistry , toxoplasma gondii , enzyme , antibody , gene , genetics , immunology
Expressed proteins and antigens from Neospora caninum tachyzoites were studied by two‐dimensional gel electrophoresis and immunoblot analysis combined with matrix‐assisted laser desorption/ionization‐time of flight mass spectrometry. Thirty‐one spots corresponding to 20 different proteins were identified from N. caninum tachyzoites by peptide mass fingerprinting. Six proteins were identified from a N. caninum database (NTPase, 14‐3‐3 protein homologue, NcMIC1, NCDG1, NcGRA1 and NcGRA2), and 11 proteins were identified in closely related species using the T. gondii database (HSP70, HSP60, pyruvate kinase, tubulin α‐ and β‐chain, putative protein disulfide isomerase, enolase, actin, fructose‐1,6‐bisphosphatase, lactate dehydrogenase and glyceradehyde‐3‐phosphate dehydrogenase). One hundred and two antigen spots were observed using pH 4–7 IPG strips on immunoblot profiles. Among them, 17 spots corresponding to 11 antigenic proteins were identified from a N. caninum protein map. This study involved the construction of in‐depth protein maps for N. caninum tachyzoites, which will be of value for studies of its pathogenesis, drug and vaccine development, and phylogenetic studies.