{"created":"2023-06-19T11:41:47.939509+00:00","id":10719,"links":{},"metadata":{"_buckets":{"deposit":"7af67a27-3b25-4f0e-932d-526f76c90883"},"_deposit":{"created_by":15,"id":"10719","owners":[15],"pid":{"revision_id":0,"type":"depid","value":"10719"},"status":"published"},"_oai":{"id":"oai:mie-u.repo.nii.ac.jp:00010719","sets":["420:724:1692754932494"]},"author_link":[],"item_8_biblio_info_6":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2007-05-01","bibliographicIssueDateType":"Issued"}}]},"item_8_description_14":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_description":"application/pdf","subitem_description_type":"Other"}]},"item_8_description_4":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"血管平滑筋は分化の進んだ筋線維の発達した「収縮型」、および未分化で増殖をする「合成型」の2種類の細胞に形質変換する。培養血管平滑筋細胞をPDGF処理によって合成型様細胞、酪酸ナトリウム(Na-butyrate, NaB)処理によって収縮型様細胞を作製し、それらの細胞に特異的な遺伝子のサブトラクションライブラリーを作成することによりスクリーニングをした。PDGF特異的ライブラリーからは230クローンが、NaB特異的ライブラリーからは160クローンの遺伝子が特定できた。これらのクローンをナイロン膜にドットブロットした膜を4枚ずつ用意し、特異性を確認するため仮想的Northernハイブリダイゼーションをおこなった。使用したプローブは、サブトラクションした2種類のcDNA、およびサブトラクション前の2種類のcDNAである。その結果有意に特異性が認められたものがPDGF処理細胞では15クローン、NaB処理細胞では19クローンに集約された。そこでこれらのクローンに対してプライマーを作製し、リアルタイムPCRをおこなって処理細胞とNaB処理細胞での発現量に有意に差があるかを確認した。2種類の細胞で発現量の差が2倍以上認められたものはCa²⁺結合蛋白質であるS100A4,S100A6、およびα-actinなどであった。そこでS100A4,S100A6の全長をクローニングし、大腸菌で発現蛋白質を作成した。さらにこの発現蛋白質とCA²⁺依存的に相互作用する因子3種類を培養平滑筋細胞から単離することができた。これらについては現在アミノ酸シークエンスを決定する準備を行っている。今後は有意に発現量に差のあった残りのクローンについても同様な解析を続けていく予定である。","subitem_description_type":"Abstract"},{"subitem_description":"Vascular smoooth muscle cell (VSMC) shows two typical phenotype, contraction-type and synthetic-type upon various situations. We developed model system of phenotypic modulation by using cultured VSMC, P53MAC01. Upon addition of PDGF, the VSMC changed to synthetic-type phenotype, and it changed to typical contraction-type phenotype in the presence of sodium butyrate (Na-B), which induce growth suppression by inhibiting acetylation of DNA. We isolated mRNA from these typical two types of cell and constructed subtraction cDNA library. We sequenced inserts of the library, and screened 230 clones from PDGF-specific library, and 160 clones from NaB-specific library. Further inserts of each clone were spotted to nylon membrane and examined by putative Northern hybridization to confirm the specificity in each types of cell. Four sets of membrane was hybridized with four sets of probes, two subtracted cDNAs and two original cDNAs from PDGF-and NaB-treated cells. As a result, 15 clones from PDGF-specific library and 19 clones from Nab-specific library were confirmed in the specificity of the expression. To quantify the expression level of these clones in PDGF-and NaB-treated cell, we made primer sets for each clone and examined with real time PCR. Clones showing 2-fold difference of expression among these two types of cell were S100A4, S100A6, a-actin and others. We isolated the full-length cDNA of S100A4 and S100A6 by RT-PCR, and expressed these genes in bacteria. Using these gene products we affinity purified three proteins that interact with S1004 and S100A6 proteins from the VSMC.","subitem_description_type":"Abstract"}]},"item_8_description_5":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"平成17年度～平成18年度科学研究費補助金(基盤研究(C))研究結果報告書","subitem_description_type":"Other"},{"subitem_description":"津","subitem_description_type":"Other"}]},"item_8_description_62":{"attribute_name":"目次","attribute_value_mlt":[{"subitem_description":"A calmodulin-dependent protein kinase from lower eukaryote Physarum polycephalum","subitem_description_type":"Other"},{"subitem_description":"Biochemical Properties of Ordinary and Dark Muscle Myosin from Carp Skeletal Muscle","subitem_description_type":"Other"},{"subitem_description":"Blebbistatin Inhibits Sphingosylphosphorylcholine-Induced Contraction of Collagen-Gel Fiber Populated by Vascular Smooth-Muscle Cells","subitem_description_type":"Other"},{"subitem_description":"Intracellular signal transduction for migration and actin remodeling in vascular smooth muscle cells after sphingosylphosphorylcholine stimulation","subitem_description_type":"Other"},{"subitem_description":"Calcium inhibition of physarum myosin as examined by the recombinant heavy mero-myosin","subitem_description_type":"Other"},{"subitem_description":"Insight Into the Mechanism of Fast Movement of Myosin From Chara corallina","subitem_description_type":"Other"},{"subitem_description":"Mechanochemical properties of ordinary and dark muscle myosins from seawater fish","subitem_description_type":"Other"},{"subitem_description":"C-protein (MyBP-C) isoforms from carp ordinary and dark muscles and muscle type-specific binding to myosin","subitem_description_type":"Other"}]},"item_8_description_64":{"attribute_name":"科研費番号","attribute_value_mlt":[{"subitem_description":"17590218","subitem_description_type":"Other"}]},"item_8_publisher_30":{"attribute_name":"出版者","attribute_value_mlt":[{"subitem_publisher":"岡垣壮"}]},"item_8_relation_46":{"attribute_name":"部分を持つ","attribute_value_mlt":[{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8660","subitem_relation_type_select":"URI"}},{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8661","subitem_relation_type_select":"URI"}},{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8662","subitem_relation_type_select":"URI"}},{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8663","subitem_relation_type_select":"URI"}},{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8802","subitem_relation_type_select":"URI"}},{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8664","subitem_relation_type_select":"URI"}},{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8665","subitem_relation_type_select":"URI"}},{"subitem_relation_type":"hasPart","subitem_relation_type_id":{"subitem_relation_type_id_text":"http://hdl.handle.net/10076/8666","subitem_relation_type_select":"URI"}}]},"item_8_subject_16":{"attribute_name":"日本十進分類法","attribute_value_mlt":[{"subitem_subject":"460","subitem_subject_scheme":"NDC"}]},"item_8_text_65":{"attribute_name":"資源タイプ（三重大）","attribute_value_mlt":[{"subitem_text_value":"Kaken / 科研費報告書"}]},"item_8_version_type_15":{"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":"岡垣, 壮","creatorNameLang":"ja"}]},{"creatorNames":[{"creatorName":"大井, 敦史","creatorNameLang":"ja"}]},{"creatorNames":[{"creatorName":"中村, 彰男","creatorNameLang":"ja"}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2017-02-20"}],"displaytype":"detail","filename":"50K5855.pdf","filesize":[{"value":"696.0 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"50K5855.pdf","url":"https://mie-u.repo.nii.ac.jp/record/10719/files/50K5855.pdf"},"version_id":"5da5956c-c3b7-4b5f-bb09-372fa04e28e6"}]},"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":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"血管平滑筋の形質変換時における筋線維の動的変化に関与する因子の検索","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"血管平滑筋の形質変換時における筋線維の動的変化に関与する因子の検索","subitem_title_language":"ja"}]},"item_type_id":"8","owner":"15","path":["1692754932494"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2008-03-18"},"publish_date":"2008-03-18","publish_status":"0","recid":"10719","relation_version_is_last":true,"title":["血管平滑筋の形質変換時における筋線維の動的変化に関与する因子の検索"],"weko_creator_id":"15","weko_shared_id":-1},"updated":"2023-09-01T01:45:30.368795+00:00"}