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相关概念视频

Proofreading01:43

Proofreading

61.9K
Overview
61.9K
Proofreading01:31

Proofreading

9.7K
Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore,  it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
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Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

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Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
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Viral Mutations00:36

Viral Mutations

40.6K
A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material...
40.6K
Bacterial Transcription01:53

Bacterial Transcription

38.5K
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:
38.5K

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Updated: Mar 19, 2026

Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis

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校对反转录酶的合成进化起源

Jared W Ellefson1, Jimmy Gollihar2, Raghav Shroff2

  • 1Center for Systems and Synthetic Biology, Institute for Cellular and Molecular Biology, Department of Molecular Biosciences, University of Texas, 2500 Speedway, Austin, TX 78712, USA. jaredwellefson@gmail.com ellingtonlab@gmail.com.

Science (New York, N.Y.)
|June 25, 2016
PubMed
概括
此摘要是机器生成的。

科学家开发了一种新的酶,反转录异聚合酶 (RTX),可以准确地将RNA复制成DNA. 这种校对酶克服了传统的逆转录酶的局限性, 实现了先进的分子生物学应用.

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Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
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科学领域:

  • 分子生物学
  • 酵素学
  • 进化生物学

背景情况:

  • 大多数逆转录酶 (RT) 缺乏校对域,导致错误率高.
  • 这种缺乏忠实性是分子生物学应用的重要局限性.

研究的目的:

  • 调查校对是否与反转录相兼容.
  • 开发一种高可靠性DNA聚合酶,能够有效地模拟RNA.

主要方法:

  • 一个热稳定DNA聚合酶的定向进化.
  • 在DNA和RNA模板上测试工程酶的活性.
  • 使用DNA和RNA基质评估校对能力.

主要成果:

  • 成功开发了一种新型酶,即逆转录异聚合酶 (RTX).
  • RTX在DNA和RNA模板上都表现出积极的校对,显著提高了准确性.
  • RTX可用于直接RNA测序和单酶RT-PCR等新型应用.

结论:

  • 校对与反向转录是一致的.
  • 工程RTX酶克服了RT忠实性的历史限制.
  • RTX为敏感和精确的分子诊断和研究开辟了新的途径.