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Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Updated: May 19, 2026

Spatiotemporally Controlled Nuclear Translocation of Guests in Living Cells Using Caged Molecular Glues as Photoactivatable Tags
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Published on: January 17, 2019

生細胞イメージングのための第2世代の共性TMPタグ.

Zhixing Chen1, Chaoran Jing, Sarah S Gallagher

  • 1Department of Chemistry, Columbia University, New York, New York 10027, USA.

Journal of the American Chemical Society
|August 10, 2012
PubMed
まとめ
この要約は機械生成です。

新しく改良されたTMPタグは,先進的な有機フッ素素を用いて,生細胞内の細胞内タンパク質のより迅速な共性ラベリングを可能にします. このブレークスルーは,生物学的研究と合成生物学アプリケーションのためのタンパク質ラベリング技術を強化します.

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Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

関連する実験動画

Last Updated: May 19, 2026

Spatiotemporally Controlled Nuclear Translocation of Guests in Living Cells Using Caged Molecular Glues as Photoactivatable Tags
10:10

Spatiotemporally Controlled Nuclear Translocation of Guests in Living Cells Using Caged Molecular Glues as Photoactivatable Tags

Published on: January 17, 2019

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10:41

Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity

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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

科学分野:

  • バイオケミストリー バイオケミストリー
  • 化学生物学 化学生物学とは
  • 合成生物学 合成生物学とは

背景:

  • オーガニック・フルオロフォアは,細胞のラベル付けのために光タンパク質よりも利点があります.
  • 以前のTMPタグの設計は,生細胞のラベリング速度と効率に制限がありました.

研究 の 目的:

  • 強化されたラベリング運動性を備えた第2世代の共振型TMPタグを開発する.
  • 生体細胞内の様々な細胞内タンパク質の効率的なラベル付けを可能にします.

主な方法:

  • アクリルアミド・トリメトプリム・フローロフォア (A-TMP-フローロフォア v2.0) を最適化されたリンク器で設計しました.
  • Escherichia coliの二水酸化葉酸還元酵素 (eDHFR) の結合ポケットの近くで設計されたシステイン (Cys) 核フィルを利用しました.
  • eDHFR:Cysの変異をスクリーニングし,迅速な共電性ラベリングのための最適なタグ・ヌクレオフィルペアを特定しました.

主要な成果:

  • 特定されたeDHFR:L28C変異種で,8分ラベル半減期 in vitro.
  • A-TMP-fluorophore v2.0プローブを使用して,様々なタンパク質標的の生細胞イメージングを成功裏に実証しました.
  • A-TMP-probe v2.0ラベルを生産するための効率的な化学合成を開発しました.

結論:

  • 第2世代の共振型TMPタグは,細胞内タンパク質のラベル付けを活体細胞に強固かつ効率的な方法として提供しています.
  • 接近誘発反応性と有機化学は,化学タグの設計と合成生物学の進歩のための強力なツールです.