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ほ乳類細胞導入直後のレポーター遺伝子に由来するmRNA量の急激な変動をリアルタイムRT-PCRにより精密に定量する方法
https://repo.qst.go.jp/records/67949
https://repo.qst.go.jp/records/67949e842d944-91ec-4f14-b5c2-15cc2f0591e3
| Item type | 会議発表用資料 / Presentation(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2004-11-29 | |||||
| タイトル | ||||||
| タイトル | ほ乳類細胞導入直後のレポーター遺伝子に由来するmRNA量の急激な変動をリアルタイムRT-PCRにより精密に定量する方法 | |||||
| 言語 | ||||||
| 言語 | jpn | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_c94f | |||||
| 資源タイプ | conference object | |||||
| アクセス権 | ||||||
| アクセス権 | metadata only access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
| 著者 |
石原, 弘
× 石原, 弘× 田中, 泉× 石原, 文子× 鈴木, 桂子× 吉野, 智恵子× 石原 弘× 田中 泉× 石原 文子× 鈴木 桂子× 吉野 智恵子 |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | Reporter gene assay is a standard method to reveal nucleotide sequence for transcriptional regulation <I>in vivo</I>. Since the activity to generate mRNA by transcriptional factors is indirectly measured from the translation product by the method, detailed function such as immediate-early response is hardly to be analyzed. Here we show a high sensitive and accurate method to analyze dynamics of reporter mRNA levels in mammalian cells. Although coexistence of large excess related DNA/RNAs, a faint level of reporter mRNAs was specifically measured by real-time RT PCR using following improvements; A reporter gene of interest was cotransfected with a reference plasmid and relative promoter level was determined. An unique sequence was attached at 5'-end of cDNA from polyA+ RNAs during reverse transcription. For the purpose, high specific primer/probe set for exogenous genes were designed. A reporter gene controlled by murine <I>jun</I>B promoter for immediate-early and short-term induction was used. After phorbol ester treatment following to transient introduction of the reporter, drastic increase and subsequent decrease of transcription were proved. The profile is closely related to mRNA levels of endogenous <I>jun</I>B gene, though a conventional reporter assay shows moderate and continuous increase . The reporter RNA assay makes an advance to analyze repression activity of nucleotide sequence <I>in vivo</I>. |
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| 会議概要（会議名, 開催地, 会期, 主催者等） | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | 第77回日本生化学会大会 | |||||
| 発表年月日 | ||||||
| 日付 | 2004-10-16 | |||||
| 日付タイプ | Issued | |||||
