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相关概念视频

Maxam-Gilbert Sequencing01:05

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In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
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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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序列特定的2 - 二醇结合

Carlo P Ramil1, Peng An1, Zhipeng Yu1

  • 1Department of Chemistry, State University of New York at Buffalo , Buffalo, New York 14260, United States.

Journal of the American Chemical Society
|April 16, 2016
PubMed
概括
此摘要是机器生成的。

研究人员发现了一种新的标签CX10R7,该标签可以在细胞环境中使用2-二 (CBT) 进行高效和特定的蛋白质标签. 这种方法最大限度地减少了蛋白质功能的干扰,为生物研究提供了一种简化方法.

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科学领域:

  • 化学生物学
  • 分子生物学
  • 生物技术

背景情况:

  • 蛋白质标签对于研究生物过程至关重要.
  • 目前的方法通常涉及恶劣的条件或复杂的催化剂,可能会扰乱蛋白质功能.
  • 需要有效的,无催化剂的生物对等反应来进行细胞内蛋白质标记.

研究的目的:

  • 发现一种用于特定区域的蛋白质标签的新标签.
  • 开发一种无催化剂的生物对角反应,用于细胞环境中的标记蛋白质.
  • 描述新发现的标签的反应性和特异性.

主要方法:

  • 使用菌素辅助反应性检测方法选一种囊编码的菌体库.
  • 设计一种名为CX10R7的2索 (CBT) 反应性标签.
  • 将CX10R7与感兴趣的蛋白质融合,并在体外和大肠杆菌细胞表面进行特定标记.
  • 进行突变发生研究以了解影响标记反应性和特异性的因素.

主要成果:

  • 发现和表征CX10R7标记,该标记与2-二 (CBT) 反应.
  • 在体外和大肠杆菌表面使用CX10R7融合蛋白进行特定位点的蛋白质标记的演示.
  • 确定周围的氨基酸序列对于稳定结合产品并确保反应性和特异性至关重要.

结论:

  • CX10R7标签提供了一种新的,高效的,无催化剂的生物对等蛋白质标记方法.
  • 这种方法最大限度地减少了蛋白质功能的干扰,使其适用于细胞应用.
  • 这些发现有助于开发蛋白质工程和生物化学研究的工具.