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Updated: May 12, 2026

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Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
Published on: April 14, 2015
基因编码的多谱标记蛋白质与多光体在DNA骨干上的多光体
Vijay Singh1, Shenliang Wang, Ke Min Chan
1Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.
Journal of the American Chemical Society
|April 18, 2013
概括
研究人员使用HaloTag技术开发了新的类似DNA的光染料,用于基因编码的蛋白质标记. 这些Oligodeoxyfluorosides (ODFs) 能够在体外和活细胞中进行高效,明亮和丰富多彩的蛋白质标记.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 化学生物学 化学生物学
背景情况:
- 遗传编码的蛋白质结合对于生物研究工具至关重要.
- 现有的蛋白质标签方法在多功能性和效率方面存在局限性.
研究的目的:
- 开发一种用于基因编码蛋白质标记的新型光染料.
- 使用HaloTag方法扩大蛋白质报告和检测能力.
主要方法:
- 开发了Oligodeoxyfluorosides (ODFs),一种具有光染色体的DNA类寡合体.
- 用于自动化染料生产的甲链接剂胺酸的合成.
- 在体外和哺乳动物细胞中,ODFs对HaloTag标记蛋白质的酶性自我结合.
主要成果:
- 通过使用九种不同的ODF染料成功地快速有效地对蛋白质进行共价标记.
- 观察到一些ODF染料在结合后的亮度增加和颜色变化.
- 在活细胞成像的各种细胞区 (细胞表面,细胞质) 中证明了ODF染料的功能.
结论:
- ODFs代表了一种通用的新类染料,用于基因编码蛋白质标签.
- 这项技术增强了活细胞中的蛋白质检测和局部化研究.
- 开发的染料为生物应用提供了更好的亮度和可调节的光谱特性.
相关概念视频
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...
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...
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...
Protein Dynamics in Living Cells
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

