関連する実験動画
Updated: Jul 26, 2026
09:28
In Vitro Transcription Assays and Their Application in Drug Discovery
Published on: September 20, 2016
トランスクリプション延長複合体の構造 in vivo
1Section of Biochemistry, Molecular, and Cell Biology, Cornell University, Ithaca, NY 14853.
Science (New York, N.Y.)
|February 14, 1992
まとめ
研究者らは,Escherichia coliの転写の延長中にDNAの開きを,一時停止部位で観察した. 彼らは,パタシウムパーマンガネートを使用して単一鎖DNAを特定し,約12個のヌクレオチドのRNA-DNAハイブリッドを明らかにしました.
科学分野:
- 分子生物学は分子生物学である.
- 遺伝学 遺伝学とは
- バイオケミストリー バイオケミストリー
背景:
- トランスクリプションは,RNAポリメラーゼを含む基本的な生物学的プロセスです.
- トランスクリプション延長中のDNA開口を理解することは,遺伝子調節に不可欠です.
研究 の 目的:
- Escherichia coliの転写延長中にインビボのDNA開口メカニズムを調査する.
- 規制停止部位における転写複合体の構造を特徴付ける.
主な方法:
- カリウムパーマンガン酸 (KMnO4) の反応性を用いて単一鎖DNAの検出.
- 転写複合体の in vivo 解析と in vitro モデルとの比較.
主要な成果:
- デュプレックスDNA開口は,レギュレータの休止部位での転写延長中に,in vivoで検出されました.
- ポタシウムパーマンガネート反応性は,転写バブル内の単一鎖DNAを特定しました.
- in vivoの構造は,in vitroの転写複合体と比較して類似点と違いを示した.
- テンプレート鎖の反応性は,約12個のヌクレオチドのRNA-DNAハイブリッドを示した.
結論:
- Escherichia coliのRNAポリメラーゼは,特定の規制部位での転写延長中にデュプレックスDNAを開く.
- RNA-DNAハイブリッドの存在は,これらの転写複合体の重要な特徴である.
関連する概念動画
02:35Transcription Elongation Factors
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
01:53Bacterial Transcription
RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
01:47Transcription Initiation
Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.
The promoters and enhancers and their accessory proteins allow tight regulation of...
The promoters and enhancers and their accessory proteins allow tight regulation of...
01:30General Transcription Factors
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
02:35Transcription Elongation Factors
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
01:28Transcription in Prokaryotes
Transcription is a highly regulated process that converts genetic information into RNA molecules. The transcription cycle is divided into three key stages: initiation, elongation, and termination, each driven by specific molecular mechanisms.Initiation of TranscriptionIn bacteria, transcription begins when the RNA polymerase core enzyme associates with a sigma factor to form a holoenzyme. For example, the E. coli sigma factor called σ70 forms a holoenzyme, which recognizes the -10 (Pribnow box)...