@misc{oai:repo.qst.go.jp:00067949,
 author = {石原, 弘 and 田中, 泉 and 石原, 文子 and 鈴木, 桂子 and 吉野, 智恵子 and 石原 弘 and 田中 泉 and 石原 文子 and 鈴木 桂子 and 吉野 智恵子},
 month = {Oct},
 note = {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>., 第77回日本生化学会大会},
 title = {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},
 year = {2004}
}