{"created":"2023-05-15T09:41:18.459891+00:00","id":402,"links":{},"metadata":{"_buckets":{"deposit":"11ae7786-983b-4541-b76e-1e8fcd8543a2"},"_deposit":{"created_by":11,"id":"402","owners":[11],"pid":{"revision_id":0,"type":"depid","value":"402"},"status":"published"},"_oai":{"id":"oai:kagawa-u.repo.nii.ac.jp:00000402","sets":["13:40"]},"author_link":["2847"],"item_10006_date_granted_11":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"2019-07-17"}]},"item_10006_degree_grantor_9":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_language":"ja","subitem_degreegrantor_name":"香川大学"},{"subitem_degreegrantor_language":"en","subitem_degreegrantor_name":"Kagawa University"}],"subitem_degreegrantor_identifier":[{"subitem_degreegrantor_identifier_name":"16201","subitem_degreegrantor_identifier_scheme":"kakenhi"}]}]},"item_10006_degree_name_8":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士（医学）","subitem_degreename_language":"ja"}]},"item_10006_description_7":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Abstract\nMarkerless gene deletion is necessary for multiple gene disruptions due to the limited number of antibiotic resistant markers for some bacteria. However, even in transformable strains, obtaining the expected mutation without a marker requires laborious screening of a large number of colonies. Previous studies had success in various bacteria with a counter-selection system where a conditional lethal gene was incorporated into the vector. We examined the efficacy of the mutated pheS gene (pheS*) as a counter-selective marker for gene deletion in Bacteroides. This mutation produces an amino acid substitution (A303G) in the alpha subunit of Bacteroides phenylalanyl tRNA synthetase, which in E. coli alters the specificity of the tRNA synthetase resulting in a conditional lethal mutation due to the incorporation of p-chloro-phenylalanine (p-Cl-Phe) into protein. B. fragilis YCH46 and B. thetaiotaomicron VPI-5482 transformed with a pheS*-harboring shuttle vector were clearly growth-inhibited in the presence of >5 mM p-Cl-Phe in liquid defined minimal media (DMM) and on DMM agar plates. A targeting plasmid was constructed to delete the genetic region for capsular polysaccharide PS2 in B. fragilis or PS1 in B. thetaiotaomicron. After counterselection, p-Cl-Phe-resistant colonies were generated at a frequency of 8.1 × 10−3 for B. fragilis and 1.7 × 10−3 for B. thetaiotaomicron. Of the p-Cl-Phe-resistant colonies, 4.2% and 72% harbored the correct genetic deletion for B. fragilis and B. thetaiotaomicron, respectively. These results indicate that mutated pheS is a useful counter-selective gene to construct markerless genetic deletions in Bacteroides.","subitem_description_language":"en","subitem_description_type":"Abstract"}]},"item_10006_dissertation_number_12":{"attribute_name":"学位授与番号","attribute_value_mlt":[{"subitem_dissertationnumber":"乙第289号"}]},"item_10006_relation_15":{"attribute_name":"関連サイト","attribute_value_mlt":[{"subitem_relation_type":"isVersionOf","subitem_relation_type_id":{"subitem_relation_type_id_text":"https://doi.org/10.1016/j.anaerobe.2016.09.004","subitem_relation_type_select":"DOI"}},{"subitem_relation_type":"isIdenticalTo","subitem_relation_type_id":{"subitem_relation_type_id_text":"https://dl.ndl.go.jp/pid/11344486","subitem_relation_type_select":"URI"}}]},"item_10006_relation_22":{"attribute_name":"PubMed番号","attribute_value_mlt":[{"subitem_relation_type_id":{"subitem_relation_type_id_text":"27639596","subitem_relation_type_select":"PMID"}}]},"item_10006_relation_28":{"attribute_name":"論文ID（NAID）","attribute_value_mlt":[{"subitem_relation_type_id":{"subitem_relation_type_id_text":"500001359041","subitem_relation_type_select":"NAID"}}]},"item_10006_rights_13":{"attribute_name":"権利","attribute_value_mlt":[{"subitem_rights":"Copyright © 2016 Elsevier Ltd. All rights reserved.","subitem_rights_language":"en"},{"subitem_rights":"この博士論文の本文については、次のエルゼビアの著作権ポリシーの規定により公開しています。「博士論文の場合は、エンバーゴ期間に関係なく機関リポジトリに出版社版を公開することができます。」 ","subitem_rights_language":"ja"},{"subitem_rights":"The text of this doctoral dissertation is published according to the following Elsevier copyright policy. \"In the case of doctoral dissertations, publishers can be published in institutional repositories regardless of the Embargo period.\"","subitem_rights_language":"en"}]},"item_10006_text_30":{"attribute_name":"KEID","attribute_value_mlt":[{"subitem_text_value":"28766"}]},"item_10006_version_type_18":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_fa2ee174bc00049f","subitem_version_type":"P"}]},"item_access_right":{"attribute_name":"アクセス権","attribute_value_mlt":[{"subitem_access_right":"open access","subitem_access_right_uri":"http://purl.org/coar/access_right/c_abf2"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorAffiliations":[{"affiliationNameIdentifiers":[{"affiliationNameIdentifier":"","affiliationNameIdentifierScheme":"ISNI","affiliationNameIdentifierURI":"http://www.isni.org/isni/"}],"affiliationNames":[{"affiliationName":"","affiliationNameLang":"ja"}]}],"creatorNames":[{"creatorName":"Kino, Yasuhiro","creatorNameLang":"en"},{"creatorName":"木納, 康博","creatorNameLang":"ja"},{"creatorName":"キノウ, ヤスヒロ","creatorNameLang":"ja-Kana"}],"familyNames":[{},{},{}],"givenNames":[{},{},{}],"nameIdentifiers":[{},{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2020-11-09"}],"displaytype":"detail","filename":"Med_B289.pdf","filesize":[{"value":"5.1 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"本文","objectType":"fulltext","url":"https://kagawa-u.repo.nii.ac.jp/record/402/files/Med_B289.pdf"},"version_id":"fe95f0e9-bf2f-4924-b59c-683fd9a3175e"},{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2020-11-09"}],"displaytype":"detail","filename":"Med_B289_abstract.pdf","filesize":[{"value":"112.7 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"内容の要旨","objectType":"abstract","url":"https://kagawa-u.repo.nii.ac.jp/record/402/files/Med_B289_abstract.pdf"},"version_id":"5020e079-4f18-4a26-803a-0da6e942fe71"},{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2020-11-09"}],"displaytype":"detail","filename":"Med_B289_result.pdf","filesize":[{"value":"610.7 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"審査の結果の要旨","objectType":"other","url":"https://kagawa-u.repo.nii.ac.jp/record/402/files/Med_B289_result.pdf"},"version_id":"9741af95-7292-4c8b-b69b-3c11c066cf9e"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"Bacteroides","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"pheS","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"tRNA","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"p-chloro-phenylalanine","subitem_subject_language":"en","subitem_subject_scheme":"Other"}]},"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":"doctoral thesis","resourceuri":"http://purl.org/coar/resource_type/c_db06"}]},"item_title":"Counterselection system employing mutated pheS for markerless genetic deletion in Bacteroides species","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Counterselection system employing mutated pheS for markerless genetic deletion in Bacteroides species","subitem_title_language":"en"}]},"item_type_id":"10006","owner":"11","path":["40"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2019-07-30"},"publish_date":"2019-07-30","publish_status":"0","recid":"402","relation_version_is_last":true,"title":["Counterselection system employing mutated pheS for markerless genetic deletion in Bacteroides species"],"weko_creator_id":"11","weko_shared_id":-1},"updated":"2024-02-08T09:06:41.045836+00:00"}