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相关概念视频

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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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

第二代共价TMP标签用于活细胞成像.

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
概括
此摘要是机器生成的。

一个新的,改进的TMP标签能够使用先进的有机光剂在活细胞中更快地对细胞内蛋白进行共价标记. 这一突破增强了用于生物研究和合成生物学应用的蛋白质标记技术.

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科学领域:

  • 生物化学 生物化学
  • 化学生物学 化学生物学
  • 合成生物学 合成生物学

背景情况:

  • 有机光体在细胞标签上比光蛋白具有优势.
  • 以前的TMP标签设计在活细胞中标记速度和效率方面存在局限性.

研究的目的:

  • 开发第二代共价TMP标签,增强标签动力学.
  • 为了能够有效地标记活细胞中的各种细胞内蛋白质.

主要方法:

  • 设计了一种具有优化链接器的烯胺-三甲-光体 (A-TMP-光体v2.0).
  • 使用一种氨酸 (Cys) 核,在大肠杆菌二叶酸减少酶 (eDHFR) 结合口袋附近进行工程.
  • 选了eDHFR:Cys变体以确定最佳的标签核对,以便快速对应标签.

主要成果:

  • 在体外鉴定了eDHFR:L28C变异,标记半衰期为8分钟.
  • 通过使用A-TMP-化物v2.0探针,证明了各种蛋白质标的活细胞成像成功.
  • 开发了一种高效的化学合成,用于生产A-TMP-probe v2.0标签.

结论:

  • 第二代共价TMP标签为活细胞中细胞内蛋白质标记提供了强大而高效的方法.
  • 接近诱导反应和有机化学是工程化学标签和推进合成生物学的强大工具.