{"created":"2021-03-01T06:34:09.868708+00:00","id":28363,"links":{},"metadata":{"_buckets":{"deposit":"ce1f9700-b1fe-42eb-9fa6-79e38d2ca4d3"},"_deposit":{"id":"28363","owners":[],"pid":{"revision_id":0,"type":"depid","value":"28363"},"status":"published"},"_oai":{"id":"oai:niigata-u.repo.nii.ac.jp:00028363","sets":["453:456","485:872:1583:1596"]},"item_7_alternative_title_1":{"attribute_name":"その他のタイトル","attribute_value_mlt":[{"subitem_alternative_title":"Synthesis of High Quality Probe Using Locked Nucleic Acid (LNA) and Exploitation for ISH"}]},"item_7_biblio_info_6":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2010-08","bibliographicIssueDateType":"Issued"},"bibliographicIssueNumber":"1","bibliographicPageEnd":"39","bibliographicPageStart":"35","bibliographicVolumeNumber":"63","bibliographic_titles":[{"bibliographic_title":"新潟大学農学部研究報告"},{"bibliographic_title":"新潟大学農学部研究報告","bibliographic_titleLang":"en"}]}]},"item_7_description_4":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"高度に保存された塩基配列を有するmRNA のin situ hybridization（ISH）による特異的検出を目的として、locked nucleicacid（LNA）を利用したキメラオリゴヌクレオチドプローブを作製し、その実用性を評価した。塩基配列で90％以上の相同性を示すカブのニトリラーゼアイソフォームBrNIT-T1及びBrNIT-T2との間で相同性の低い24～25塩基の領域に相補的なオリゴヌクレオチドを設計し、両者間で異なる塩基（９～10塩基）をLNA に置換したLNA-DNA キメラオリゴヌクレオチドを作製した。配列中にLNA を部分的に導入することにより、DNA のみでは55～57℃であったオリゴヌクレオチドのTm 値は78～82℃に上昇し、ISH 解析に十分な値を示した。BrNIT-T1及びBrNIT-T2の全長転写産物に対する結合性をドットブロットノザンハイブリダイゼーションにより調査したところ、BrNIT-T1及びBrNIT-T2のそれぞれの配列に対応するLNA-DNA キメラプローブは互いの転写産物に非特異的に結合することなく、目的の転写産物のみを検出できることが確認された。さらにこれらのキメラプローブをカブ根こぶ病組織のISH 解析に応用し、従来のcRNA プローブを用いた解析では得られなかった、BrNIT-T1とBrNIT-T2の時空間的な発現特異性を明らかにすることに成功した。","subitem_description_type":"Abstract"},{"subitem_description":"To achieve specific detection of highly-conserved sequences by in situ hybridization (ISH), locked nucleic acid(LNA)-containing oligonucleotide probes were designed and assessed. Two isoforms of turnip nitrilases, BrNIT-T1 andBrNIT-T2, that possess more than 90% homology at the nucleotide level, were targeted in this study. Oligonucleotidescorresponding to a relatively different region between BrNIT-T1 and BrNIT-T2 were generated to contain LNAs in placeof DNA bases different between the two sequences. The partial incorporation of LNA into DNA oligonucleotides providedsufficient Tm values for ISH analysis. Dot-blot hybridization assay confirmed that the two LNA-modified oligonucleotideprobes corresponding to sequences of BrNIT-T1 and BrNIT-T2 specifically hybridized with the desired RNA sequencebut not with another one. Advantages of the LNA-modified probes in ISH analysis were also observed as specific andsensitive detection of BrNIT-T localization. Based on these results, we here report that partial incorporation of LNA intooligonucleotide probe is useful for highly specific detection of similar genes in ISH analysis.","subitem_description_type":"Abstract"}]},"item_7_full_name_3":{"attribute_name":"著者別名","attribute_value_mlt":[{"nameIdentifiers":[{"nameIdentifier":"163139","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Ishikawa, Toshiki"}]},{"nameIdentifiers":[{"nameIdentifier":"163140","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Okazaki, Keiichi"}]},{"nameIdentifiers":[{"nameIdentifier":"163141","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Itoh, Kimiko"}]},{"nameIdentifiers":[{"nameIdentifier":"163142","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Mitsui, Toshiaki"}]},{"nameIdentifiers":[{"nameIdentifier":"163143","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Hori, Hidetaka"}]}]},"item_7_publisher_7":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"新潟大学農学部"}]},"item_7_select_19":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_select_item":"publisher"}]},"item_7_source_id_11":{"attribute_name":"書誌レコードID","attribute_value_mlt":[{"subitem_source_identifier":"AN00183393","subitem_source_identifier_type":"NCID"}]},"item_7_source_id_9":{"attribute_name":"ISSN","attribute_value_mlt":[{"subitem_source_identifier":"03858634","subitem_source_identifier_type":"ISSN"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"石川, 寿樹"}],"nameIdentifiers":[{"nameIdentifier":"163134","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"岡崎, 圭一"}],"nameIdentifiers":[{"nameIdentifier":"163135","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"伊藤, 紀美子"}],"nameIdentifiers":[{"nameIdentifier":"163136","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"三ツ井, 敏明"}],"nameIdentifiers":[{"nameIdentifier":"163137","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"堀, 秀隆"}],"nameIdentifiers":[{"nameIdentifier":"163138","nameIdentifierScheme":"WEKO"}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2019-08-20"}],"displaytype":"detail","filename":"63(1)_35-39.pdf","filesize":[{"value":"490.9 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"63(1)_35-39.pdf","url":"https://niigata-u.repo.nii.ac.jp/record/28363/files/63(1)_35-39.pdf"},"version_id":"e2fa8c1a-f76a-4583-82a4-6512194c663c"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"locked nucleic acid (LNA)","subitem_subject_scheme":"Other"},{"subitem_subject":"in situ hybridization (ISH)","subitem_subject_scheme":"Other"},{"subitem_subject":"キメラオリゴヌクレオチドプローブ","subitem_subject_scheme":"Other"},{"subitem_subject":"ニトリラーゼ","subitem_subject_scheme":"Other"},{"subitem_subject":"Brassica rapa","subitem_subject_scheme":"Other"},{"subitem_subject":"Chimera oligonucleotide probe","subitem_subject_scheme":"Other"},{"subitem_subject":"nitrilase","subitem_subject_scheme":"Other"},{"subitem_subject":"Brassica rapa","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"departmental bulletin paper","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"Locked nucleic acid（LNA) を利用した高性能ハイブリダイゼーションプローブの作製","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Locked nucleic acid（LNA) を利用した高性能ハイブリダイゼーションプローブの作製"},{"subitem_title":"Locked nucleic acid（LNA) を利用した高性能ハイブリダイゼーションプローブの作製","subitem_title_language":"en"}]},"item_type_id":"7","owner":"1","path":["456","1596"],"pubdate":{"attribute_name":"公開日","attribute_value":"2012-07-02"},"publish_date":"2012-07-02","publish_status":"0","recid":"28363","relation_version_is_last":true,"title":["Locked nucleic acid（LNA) を利用した高性能ハイブリダイゼーションプローブの作製"],"weko_creator_id":"1","weko_shared_id":null},"updated":"2022-12-15T03:58:24.998681+00:00"}