{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T23:21:31Z","timestamp":1774567291880,"version":"3.50.1"},"reference-count":29,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2000,6,1]],"date-time":"2000-06-01T00:00:00Z","timestamp":959817600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2000,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Current serological tests for\n            <jats:italic>Babesia bigemina<\/jats:italic>\n            use semipurified merozoite antigens derived from infected erythrocytes. One of the major drawbacks of these tests is that antigen quality can vary from batch to batch. Since the quality of the antigen contributes to the sensitivity and specificity of serological tests, the use of standardized recombinant antigens should ensure consistency in assay quality. Previously, a 200-kDa merozoite antigen (p200) was identified as a candidate diagnostic antigen for use in a serological assay for the detection of\n            <jats:italic>B. bigemina<\/jats:italic>\n            antibodies in infected cattle. In this study, we have cloned, characterized, and expressed p200. A 3.5-kbp cDNA clone encoding p200 was isolated and shown to be almost full length, lacking approximately 300 bp at the 5\u2032 end. The predicted amino acid sequence shows that p200 consists of a long, highly charged central repeat region of an uninterrupted \u03b1 helix, indicative of a fibrous protein. Immunoelectron microscopy localized p200 to the merozoite cytoplasm, suggesting that the antigen may be a structural protein involved in forming filament structures within the cytoskeleton. The 3.5-kbp cDNA was expressed in bacteria as a fusion protein with glutathione\n            <jats:italic>S<\/jats:italic>\n            -transferase (GST), but the yield was poor. To improve the yield, cDNA fragments encoding antigenic domains of p200 were expressed as fusions with GST. One of these fusion proteins, C1A-GST, is composed of a 7-kDa fragment of the p200 repeat region and contains epitopes that react strongly with sera from cattle experimentally infected with\n            <jats:italic>B. bigemina<\/jats:italic>\n            . Recombinant C1A-GST should permit the development of an improved enzyme-linked immunosorbent assay for the detection of antibodies against\n            <jats:italic>B. bigemina.<\/jats:italic>\n          <\/jats:p>","DOI":"10.1128\/jcm.38.6.2240-2247.2000","type":"journal-article","created":{"date-parts":[[2020,3,9]],"date-time":"2020-03-09T19:47:19Z","timestamp":1583783239000},"page":"2240-2247","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":33,"title":["Cloning, Characterization, and Expression of a 200-Kilodalton Diagnostic Antigen of\n            <i>Babesia bigemina<\/i>"],"prefix":"10.1128","volume":"38","author":[{"given":"N.","family":"Tebele","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->International Livestock Research Institute, Nairobi, Kenya,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. A.","family":"Skilton","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->International Livestock Research Institute, Nairobi, Kenya,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J.","family":"Katende","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->International Livestock Research Institute, Nairobi, Kenya,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C. W.","family":"Wells","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->International Livestock Research Institute, Nairobi, Kenya,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"V.","family":"Nene","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->International Livestock Research Institute, Nairobi, Kenya,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T.","family":"McElwain","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington2"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S. P.","family":"Morzaria","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->International Livestock Research Institute, Nairobi, Kenya,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A. J.","family":"Musoke","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->International Livestock Research Institute, Nairobi, Kenya,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1177\/38.2.1688894"},{"key":"e_1_3_2_3_2","unstructured":"Brown\nC. G. D.\nHunter\nA. G.\nLuckins\nA. G.\nDiseases caused by protozoa\nHandbook on animal diseases in the tropics\n4th ed.\nSewell\nM. M. H.\nBrocklesby\nD. W.\n1990\n161\n226\nBailli\u00e8re Tindall\nLondon England"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.120.2.339"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.34.11.2748-2755.1996"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.86.6.1885"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(95)00036-Z"},{"key":"e_1_3_2_8_2","unstructured":"de Vos\nA. J.\nJorgensen\nW. K.\nBovine babesiosis\nManual of recommended diagnostic techniques and requirements for biological products.\n1991\n1\/14\n14\/14\nOffice International des Epizootics\nParis France"},{"key":"e_1_3_2_9_2","unstructured":"de Vos\nA. J.\nPotgieter\nF. T.\nDe Waal\nD. T.\nvan Heerden\nJ.\nBabesioses\nInfectious diseases of livestock with special reference to southern Africa.\nCoetzer\nJ. A. W.\nThompson\nG. R.\nTustin\nR. C.\n1994\n278\n308\nOxford University Press\nOxford United Kingdom"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(78)90297-8"},{"key":"e_1_3_2_11_2","unstructured":"Hames\nB. D.\nOne-dimensional polyacrylamide gel electrophoresis\nGel electrophoresis of proteins. A practical approach.\nHames\nB. D.\nRickwood\nD.\n1990\n1\n147\nOxford University Press\nOxford United Kingdom"},{"key":"e_1_3_2_12_2","unstructured":"Harlow\nE.\nLane\nD.\nAntibodies: a laboratory manual.\n1988\nCold Spring Harbor Laboratory Press\nCold Spring Harbor N.Y"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/277491a0"},{"key":"e_1_3_2_14_2","unstructured":"Jorgensen\nW.\nde Vos\nA. J.\nDalgleish\nJ. R.\nMethods for diagnosis of babesiosis in developing countries. Present and future trends\nUse of applicable biotechnological methods for diagnosing haemoparasites.\nUilenberg\nG.\nPermin\nA.\nHansen\nJ. W.\n1994\n65\n84\nFAO\nRome Italy"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004360050419"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.mi.41.100187.001145"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(92)90446-V"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/0304-4017(81)90061-3"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1590\/S0074-02761992000700033"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(80)90168-4"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/21.11.2783"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1006\/mcpr.1998.0223"},{"key":"e_1_3_2_23_2","unstructured":"Sambrook\nJ.\nFritsch\nE. F.\nManiatis\nT.\nMolecular cloning: a laboratory manual\n2nd ed.\n1989\nCold Spring Harbor Laboratory Press\nCold Spring Harbor N.Y"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(89)91995-5"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(88)90005-4"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(84)90029-X"},{"key":"e_1_3_2_27_2","unstructured":"Tebele\nN.\nPh.D. thesis.\n1996\n Brunel University\nUxbridge United Kingdom"},{"key":"e_1_3_2_28_2","doi-asserted-by":"crossref","unstructured":"Weiland\nG.\nReiter\nI.\nMethods for the measurement of the serological response to Babesia Babesiosis of domestic animals and man.\nRistic\nM.\n1988\n143\n162\nCRC Press Inc.\nBoca Raton Fla","DOI":"10.1201\/9781351070027-9"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.20506\/rst.9.2.508"},{"key":"e_1_3_2_30_2","unstructured":"Zubay\nG.\nMethods for characterization and purification of proteins\nBiochemistry.\n1988\n100\n130\nMacMillan Publishing Company\nNew York N.Y"}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.38.6.2240-2247.2000","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.38.6.2240-2247.2000","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,5]],"date-time":"2022-03-05T00:41:48Z","timestamp":1646440908000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JCM.38.6.2240-2247.2000"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2000,6]]},"references-count":29,"aliases":["10.1128\/.38.6.2240-2247.2000"],"journal-issue":{"issue":"6","published-print":{"date-parts":[[2000,6]]}},"alternative-id":["10.1128\/JCM.38.6.2240-2247.2000"],"URL":"https:\/\/doi.org\/10.1128\/jcm.38.6.2240-2247.2000","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2000,6]]},"assertion":[{"value":"1999-09-16","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2000-03-29","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2000-06-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}