{"created":"2023-05-15T16:32:35.161724+00:00","id":4487,"links":{},"metadata":{"_buckets":{"deposit":"56a0a3b2-cb48-4471-b06f-881529d45ecf"},"_deposit":{"created_by":2,"id":"4487","owners":[2],"pid":{"revision_id":0,"type":"depid","value":"4487"},"status":"published"},"_oai":{"id":"oai:nagasaki-u.repo.nii.ac.jp:00004487","sets":["35:36"]},"author_link":["17901","17898","17905","17902","17903","17899","17900","17904"],"item_2_biblio_info_6":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2015-05-11","bibliographicIssueDateType":"Issued"},"bibliographicIssueNumber":"5","bibliographicPageStart":"e0125993","bibliographicVolumeNumber":"10","bibliographic_titles":[{"bibliographic_title":"PLOS ONE"}]}]},"item_2_description_4":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Genetic manipulation is an essential technique to analyze gene function; however, limited methods are available for Babesia bovis, a causative pathogen of the globally important cattle disease, bovine babesiosis. To date, two stable transfection systems have been developed for B. bovis, using selectable markers blasticidin-S deaminase (bsd) or human dihydrofolate reductase (hdhfr). In this work, we combine these two selectable markers in a sequential transfection system. Specifically, a parent transgenic B. bovis line which episomally expresses green fluorescent protein (GFP) and human dihydrofolate reductase (hDHFR), was transfected with a plasmid encoding a fusion protein consisting of red fluorescent protein (RFP) and blasticidin-S deaminase (BSD). Selection with WR99210 and blasticidin-S resulted in the emergence of parasites double positive for GFP and RFP. We then applied this method to complement gene function in a parasite line in which thioredoxin peroxidase-1 (Bbtpx-1) gene was knocked out using hDHFR as a selectable marker. A plasmid was constructed harboring both RFP-BSD and Bbtpx-1 expression cassettes, and transfected into a Bbtpx-1 knockout (KO) parasite. Transfectants were independently obtained by two transfection methods, episomal transfection and genome integration. Complementation of Bbtpx-1 resulted in full recovery of resistance to nitrosative stress, via the nitric oxide donor sodium nitroprusside, which was impaired in the Bbtpx-1 KO parasites. In conclusion, we developed a sequential transfection method in B. bovis and subsequently applied this technique in a gene complementation study. This method will enable broader genetic manipulation of Babesia toward enhancing our understanding of the biology of this parasite.","subitem_description_type":"Abstract"}]},"item_2_description_63":{"attribute_name":"引用","attribute_value_mlt":[{"subitem_description":"PLOS ONE, 10(5), e0125993; 2015","subitem_description_type":"Other"}]},"item_2_publisher_33":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"Public Library of Science"}]},"item_2_relation_12":{"attribute_name":"DOI","attribute_value_mlt":[{"subitem_relation_type":"isIdenticalTo","subitem_relation_type_id":{"subitem_relation_type_id_text":"10.1371/journal.pone.0125993","subitem_relation_type_select":"DOI"}}]},"item_2_rights_13":{"attribute_name":"権利","attribute_value_mlt":[{"subitem_rights":"c 2015, Public Library of Science. All rights reserved. This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication."}]},"item_2_source_id_8":{"attribute_name":"EISSN","attribute_value_mlt":[{"subitem_source_identifier":"19326203","subitem_source_identifier_type":"ISSN"}]},"item_2_version_type_16":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"Asada, Masahito"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Yahata, Kazuhide"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Hakimi, Hassan"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Yokoyama, Naoaki"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Igarashi, Ikuo"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Kaneko, Osamu"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Suarez, Carlos E."}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Kawazu, Shin-ichiro"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2020-12-21"}],"displaytype":"detail","filename":"PLoS10_125993.pdf","filesize":[{"value":"1.3 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"PLoS10_125993.pdf","url":"https://nagasaki-u.repo.nii.ac.jp/record/4487/files/PLoS10_125993.pdf"},"version_id":"0e2fc5fe-2106-478b-bccf-c708f2b11ad4"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"journal article","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"Transfection of Babesia bovis by Double Selection with WR99210 and Blasticidin-S and Its Application for Functional Analysis of Thioredoxin Peroxidase-1","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Transfection of Babesia bovis by Double Selection with WR99210 and Blasticidin-S and Its Application for Functional Analysis of Thioredoxin Peroxidase-1"}]},"item_type_id":"2","owner":"2","path":["36"],"pubdate":{"attribute_name":"公開日","attribute_value":"2015-07-09"},"publish_date":"2015-07-09","publish_status":"0","recid":"4487","relation_version_is_last":true,"title":["Transfection of Babesia bovis by Double Selection with WR99210 and Blasticidin-S and Its Application for Functional Analysis of Thioredoxin Peroxidase-1"],"weko_creator_id":"2","weko_shared_id":-1},"updated":"2023-05-16T03:20:58.164135+00:00"}