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ID 40722
フルテキストURL
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タイトル(別表記)
Detection of gene expression in cells trasfected with recombinant DNA by in situ hybridization
著者
村上 雅 岡山大学医学部癌源研究施設生化学部門
抄録
Expression of mRNA in monolayer cells transfected with recombinant DNA was demonstrated by in situ hybridization with biotin- or (32)P-labeled probes. The biotin-labeled probe and hybrid detection system used in this study could detect 1 pg of target DNA and a single copy gene in mammalian cells using filter hybridization techniques. Several conditions of in situ hybridization were optimized with neomycin resistant (neo(r)) cells, which constitutively expressed neo(r) mRNA derived from pSV2neo. The proteolytic digestion was found to be the most critical procedure. The in situ hybridization demonstrated that neo(r) mRNA was localized in the cytoplasm of the neo(r) cells. Cf2Th cells derived from a canine fetal thymus cell were transfected with the cloned provirus genome of a retrovirus produced in a human lymphoblastoid cell line, and transient expressions of viral RNA and antigens were monitored by in situ hybridization and immunoperoxidase staining, respectively. About 48 hours after transfection, several percent of the transfected cells were positive for expression of both viral RNA and antigens by these cytochemical methods. These observations indicate that in situ hybridization is a useful method for detecting the expression of the gene introduced into mammalian cells.
キーワード
in situハイブリダイゼイション
ビオチン標識プローブDNAトランスフェクション
遺伝子発現
発行日
1989-02
出版物タイトル
岡山医学会雑誌
出版物タイトル(別表記)
Journal of Okayama Medical Association
101巻
1-2号
出版者
岡山医学会
出版者(別表記)
Okayama Medical Association
開始ページ
155
終了ページ
166
ISSN
0030-1558
NCID
AN00032489
資料タイプ
学術雑誌論文
オフィシャル URL
https://www.jstage.jst.go.jp/article/joma1947/101/1-2/101_1-2_155/_article/-char/ja/
関連URL
http://www.okayama-u.ac.jp/user/oma/
言語
Japanese
著作権者
岡山医学会
論文のバージョン
publisher
査読
有り
Eprints Journal Name
joma