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相关概念视频

From DNA to Protein03:06

From DNA to Protein

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The flow of genetic information in cells from DNA to mRNA to protein is described by the central dogma, which states that genes specify the sequence of mRNAs, which in turn specify the sequence of amino acids making up all proteins. The decoding of one molecule to another is performed by specific proteins and RNAs. Because the information stored in DNA is so central to cellular function, it makes intuitive sense that the cell would make mRNA copies of this information for protein synthesis...
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The Central Dogma01:25

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Overview
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Leaky Scanning02:28

Leaky Scanning

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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...
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tRNA Activation02:26

tRNA Activation

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Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
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Initiation of Translation02:33

Initiation of Translation

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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
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Exon Recombination02:32

Exon Recombination

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The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon...
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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
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哺乳动物细胞中的遗传密码扩展通过四重解码.

Yan Chen1, Tianyu Gao1, Xinyuan He2

  • 1Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE, USA.

Methods in molecular biology (Clifton, N.J.)
|June 5, 2023
PubMed
概括

这项研究引入了一种新的方法,用于在哺乳动物细胞中使用四重编码子进行特定位点的蛋白质修饰. 这种遗传密码扩展技术允许精确插入非正规氨基酸 (ncAAs) 进行先进的生物研究.

关键词:
四个基础的编码子.遗传密码扩展 遗传密码扩展非正规氨基酸非正规氨基酸四个小的鱼.不自然的氨基酸是一种非自然的氨基酸.

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科学领域:

  • 生物化学 生物化学
  • 分子生物学分子生物学
  • 合成生物学 合成生物学

背景情况:

  • 遗传密码的扩展允许将非正规氨基酸 (ncAAs) 具体地纳入蛋白质中.
  • 扩展遗传代码超出三重代码,例如使用四重代码,为蛋白质工程提供了新的可能性.

研究的目的:

  • 在哺乳动物细胞中提供一种解码UAGA四倍编码子与ncAA的协议.
  • 用工程氨基酸-tRNA合成酶 (aaRS) 和tRNA变体来演示基因纳入ncAAs的一般方法.

主要方法:

  • 使用一种工程氨基-tRNA合成酶 (aaRS) 和一种具有扩展抗环的tRNA变体.
  • 开发一种用于哺乳动物细胞中的四重编码子解码的协议.
  • 使用显微镜成像和流细胞计进行分析.

主要成果:

  • 在哺乳动物细胞中成功解码了UAGA四倍编码子与ncAA.
  • 通过四重代码子来进行ncAAs的遗传结合的可通用方法的演示.
  • 使用先进的成像和流细胞计技术进行ncAA突变发生的表征.

结论:

  • 开发的协议使得哺乳动物细胞中的遗传代码能够使用四重代码扩展.
  • 这种方法为特定位点的蛋白质修饰和研究ncAA突变发生提供了强大的工具.
  • 该方法广泛适用于推进蛋白质工程和合成生物学应用.