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相关概念视频

Ribosome Profiling02:24

Ribosome Profiling

3.5K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.5K
Ribosomes01:27

Ribosomes

7.5K
Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that synthesize large quantities of protein—such as secretory cells in the human pancreas—can contain millions of ribosomes.
Ribosome Structure and Assembly
Ribosomes are composed of ribosomal RNA (rRNA) and proteins. In eukaryotes, rRNA is transcribed from genes in the nucleolus—a part of the nucleus that specializes in ribosome...
7.5K
Improving Translational Accuracy02:07

Improving Translational Accuracy

10.2K
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
10.2K
Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

13.2K
Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
13.2K
Leaky Scanning02:28

Leaky Scanning

5.1K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.1K
Post-translational Translocation of Proteins to the RER01:27

Post-translational Translocation of Proteins to the RER

5.7K
A sizable fraction of proteins destined for ER are first synthesized in the cell cytosol and then transported across the ER membrane–a process called post-translational translocation. Similar to cotranslationally translocated proteins, these proteins also use the Sec translocon complex to enter the ER lumen.
Targeting proteins to the ER
Hsp40 and Hsp70 chaperone molecules bind the translated proteins in the cytosol to prevent their folding. The chaperone binding helps to keep the signal...
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相关实验视频

Updated: Jun 27, 2025

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

Published on: April 27, 2018

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核糖体池工程增加了蛋白质生物合成产量.

Camila Kofman1, Jessica A Willi1, Ashty S Karim1

  • 1Department of Chemical and Biological Engineering, Northwestern University, Evanston, Illinois 60208, United States.

ACS central science
|April 29, 2024
PubMed
概括
此摘要是机器生成的。

细菌核糖体具有影响蛋白质生产的序列变化. 通过使用特定的核糖体RNA (rRNA) 操作子来设计这些核糖体,可以显著提高合成生物学和生物制造的蛋白质产量.

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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
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Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling
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Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling

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相关实验视频

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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
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科学领域:

  • 分子生物学分子生物学
  • 合成生物学 合成生物学
  • 细菌遗传学 细菌遗传学

背景情况:

  • 细菌核糖体对于蛋白质合成至关重要,具有多样化的核糖体RNA (rRNA) 操作序列.
  • 由于这些本源序列变异而产生的功能差异尚未得到充分理解.
  • 了解这些差异是优化合成生物学应用中的核糖体的关键.

研究的目的:

  • 为了研究 *Escherichia coli* rRNA操作子中的序列多态性对核糖体性能的功能影响.
  • 评估由不同rRNA操作子组合衍生的核糖体的蛋白质生产能力.
  • 探索工程核糖体池的潜力,以提高蛋白质生物制造产量.

主要方法:

  • 利用*体外*的核糖体合成和翻译平台,分析来自七个*大肠杆菌*的rRNA操作子中的16S和23SrRNA独特组合的核糖体.
  • 产生表达单个rRNA操作子的细菌菌株,用于*in vivo*分析.
  • 使用医学和工业相关蛋白质组的评估蛋白质生产产量.

主要成果:

  • 原生 *E. coli* rRNA 操作子的序列变化显著改变了核糖体功能和蛋白质生产效率.
  • 与野生类型异质池相比,一些纯化*in vivo*表达的同质核糖体池表现出优异的性能.
  • 一个仅表达操作子A的细胞溶解体的核糖体显示了对标蛋白的实质性产量增加.

结论:

  • 原生rRNA操作子序列多态性直接影响核糖体功能和蛋白质合成.
  • 通过选择特定的rRNA操作子来设计核糖体池,可以提高生物制造中的蛋白质产量.
  • 核糖体池工程为优化蛋白质生产和理解核糖体进化提供了一种新的策略.