{"created":"2023-07-27T06:50:06.100692+00:00","id":42580,"links":{},"metadata":{"_buckets":{"deposit":"a22202a9-fdaa-4bc8-bc9f-69cad0f4622a"},"_deposit":{"created_by":18,"id":"42580","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"42580"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00042580","sets":["2812:2813:2829"]},"author_link":["20455","67963"],"item_10007_biblio_info_24":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2005-04","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"4p.","bibliographicVolumeNumber":"2003-2004","bibliographic_titles":[{"bibliographic_title":"平成16(2004)年度 科学研究費補助金 基盤研究(C) 研究成果報告書"},{"bibliographic_title":"2004 Fiscal Year Final Research Report","bibliographic_titleLang":"en"}]}]},"item_10007_date_8":{"attribute_name":"報告年度","attribute_value_mlt":[{"subitem_date_issued_datetime":"2005-04","subitem_date_issued_type":"Issued"}]},"item_10007_description_13":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Y染色体のゲノム塩基配列が公開されると、相同性の高い反復配列とパリンドローム構造であることが明らかとなった。Y染色体上の大きなパリンドローム構造は8種あり、精子形成候補領域のひとつであるAZFcにはふたつのパリンドローム複合体が存在している。我々は、パリンドローム単位で欠失が生じているという仮定のもとに、無・高度乏精子症患者ゲノムDNAを用いて、AZFcパリンドローム複合体の有無について検討した。まず、現在多用されている多コピー遺伝子や反復・繰返し配列の多いAZFc内部の欠失を確認することは従来のSTS-PCR法では理論的に不可能である。したがって、ゲノム塩基配列から、それぞれのプローブに対応するコピー数をreal-time quantitative PCR (RTQ PCR)によって確認し、特発性男性不妊症との関係について検討した。まず、定量化するために内部標準DNAを鋳型にして標準曲線を作成した。この鋳型は常染色体上の単一遺伝子であるRNAase P遺伝子を使用し、各サンプルのゲノムDNA量のばらつきを補正した。各患者サンプルの鋳型DNA量を10ngとし、RTQ-PCRを40サイクル行った。RTQ-PCRより算出できるコピー数比から、パリンドローム内部における微小欠失を検出することが可能となった。特に、パリンドローム部分欠失の検出は理論上無理とされていたが、ゲノムデータベースを基礎に男性不妊症でその部分欠失の存在を証明した。AZFc領域にあるP1+P2欠失のみならず、P1部分欠失の方法を確立した。今回使用したRTQ-PCR法において、従来のSTS-PCR法では欠失を認められない症例にも、AZFc領域内部に微小欠失が存在していることが明らかとなった。従来考えられていたよりも多くの特発性男性不妊症患者にAZFc領域の微小欠失が存在している可能性が考えられた。","subitem_description_type":"Abstract"},{"subitem_description":"Background :\nIn 2003, the whole sequence of the Y chromosome has been determined following the completion of the human genome project. As a result, huge identical sequences have been found to be present massive Y-specific repeat called amplicons on the distal side of the euchromatic region of the long arm of the Y chromosome, and these amliconic regions have formed a massive palindrome complex. The palindromes in the AZFc region are consists of a complex of several small segments. These ampliconic sequences pairs are characterised by palindromes showing nearly more than 99.9% identy. Sequence tagged site (STS) markers are now available for detection of the position of palindromes. The AZFc region is comprised completely of amplicons and is particularly susceptible to deletion. The b2/b4 deletion is eliminated the entire AZFc region and cause the spermatogenetic failure. Some papers reported partial deletions within AZFc are present in some infertile male. Conventional PCR is not evaluat ed a copy number, but, real-time PCR assay that may be adaptable for estimating several identical sequence sites as copies numbers.\nThe objective of the present study was to evaluate the multiple sites which is identical to sequences using the quantitative detection of copies by real-time fluorescence PCR as a new molecular diagnostic parameter in the genome DNA from patients presenting with non-obstructive azoospermia. Here we apply quantitative real-time PCR as an alternative approach to measure DNA copy number changes at each AZF region of the human Y chromosome.\nWe selcted the probes such as sY 142 (G38345), sY 254 (G38349), sY 579 (G63909), sY 602 (G34986), sY 627 (G67175), sY 639 (G67162), sY 1054(G6716), sY 1125 (G67164), sY 1190 (G67165), sY 1192 (G67166), sY 1196 (G67167), sY 1197 (G67168), sY 1198 (G67169), sY 1201 (G67170), sY 1206 (G67171). We modified these primers size for real time PCR, so that the PCR products products is amplified approximately 100 base pairs and hybridization probes within each primer and dual-labelled flurogenic probes (hybridization probe) is designed between these selected modified primers. These amplicons and corresponding primers that showed a unique BLAST hit were selected and used in further experiments.\nThe region of AZFc are present one or more identifical sequences per genome because of their palindromic structures. As more several retetive sequence exist, the possibility of partial deletion within AZFc is present.\nWe here demonstrated the application of a new, flexible, fast, and precise real-time PCR based estimation the copy number of identical sequences in Y chromosome ampliconic region.","subitem_description_type":"Abstract"}]},"item_10007_description_14":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:15591677, 研究期間(年度):2003-2004","subitem_description_type":"Other"},{"subitem_description":"出典：「特発性男性不妊症患者に対するY染色体パリンドローム複合体の分析と多型性検索」研究成果報告書　課題番号15591677\n(KAKEN：科学研究費助成事業データベース（国立情報学研究所）)\n　　　本文データは著者版報告書より作成","subitem_description_type":"Other"}]},"item_10007_description_4":{"attribute_name":"研究代表者ID","attribute_value_mlt":[{"subitem_description":"90283134","subitem_description_type":"Other"}]},"item_10007_description_9":{"attribute_name":"研究課題番号","attribute_value_mlt":[{"subitem_description":"15591677","subitem_description_type":"Other"}]},"item_10007_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00048935","subitem_identifier_reg_type":"JaLC"}]},"item_10007_relation_17":{"attribute_name":"関連サイト","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/search/?qm=90283134"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/search/?qm=90283134","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15591677/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15591677/","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-15591677/155916772004kenkyu_seika_hokoku_gaiyo/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-15591677/155916772004kenkyu_seika_hokoku_gaiyo/","subitem_relation_type_select":"URI"}}]},"item_10007_version_type_20":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2017-11-16"}],"displaytype":"detail","filename":"ME-PR-KOH-E-kaken 2005-4p.pdf","filesize":[{"value":"59.3 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-KOH-E-kaken 2005-4p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/42580/files/ME-PR-KOH-E-kaken 2005-4p.pdf"},"version_id":"57e36c4a-d309-4afa-a05f-42ec0be3baa8"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"Y染色体","subitem_subject_scheme":"Other"},{"subitem_subject":"男性不妊症","subitem_subject_scheme":"Other"},{"subitem_subject":"AZF","subitem_subject_scheme":"Other"},{"subitem_subject":"パリンドローム","subitem_subject_scheme":"Other"},{"subitem_subject":"ゲノム","subitem_subject_scheme":"Other"},{"subitem_subject":"STS","subitem_subject_scheme":"Other"},{"subitem_subject":"real-time PCR","subitem_subject_scheme":"Other"},{"subitem_subject":"精子形成障害","subitem_subject_scheme":"Other"},{"subitem_subject":"Y chromosome","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"male infertility","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"AZF","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"palindrome","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"genome","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"STS","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"real-time PCR","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Spermatogenesis","subitem_subject_language":"en","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_researcher":{"attribute_name":"研究代表者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"高, 栄哲"}],"nameIdentifiers":[{"nameIdentifier":"67963","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"90283134","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=90283134"}]},{"creatorNames":[{"creatorName":"Koh, Eitetsu"}],"nameIdentifiers":[{"nameIdentifier":"20455","nameIdentifierScheme":"WEKO"},{"nameIdentifier":"90283134","nameIdentifierScheme":"e-Rad","nameIdentifierURI":"https://kaken.nii.ac.jp/ja/search/?qm=90283134"},{"nameIdentifier":"90283134","nameIdentifierScheme":"研究者番号","nameIdentifierURI":"https://nrid.nii.ac.jp/nrid/1000090283134"}]}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"特発性男性不妊症患者に対するY染色体パリンドローム複合体の分析と多型性検索","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"特発性男性不妊症患者に対するY染色体パリンドローム複合体の分析と多型性検索"},{"subitem_title":"Analysis and detecction of polymorphism of palindrome complex on Y chmmosome in idiopathic male infertility","subitem_title_language":"en"}]},"item_type_id":"10007","owner":"18","path":["2829"],"pubdate":{"attribute_name":"公開日","attribute_value":"2017-11-16"},"publish_date":"2017-11-16","publish_status":"0","recid":"42580","relation_version_is_last":true,"title":["特発性男性不妊症患者に対するY染色体パリンドローム複合体の分析と多型性検索"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T14:29:25.365908+00:00"}