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Related Concept Videos

Proteomics01:33

Proteomics

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A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
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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...
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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
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Protein expression systems for structural genomics and proteomics.

Shigeyuki Yokoyama1

  • 1RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045, Japan. yokoyama@biochem.s.u-tokyo.ac.jp

Current Opinion in Chemical Biology
|January 28, 2003
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Summary
This summary is machine-generated.

High-throughput protein expression for structural biology is optimized using robotic cloning and microbial hosts. Cell-free systems aid large-scale production, though protein folding remains a challenge, addressed by screening methods.

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Area of Science:

  • Structural biology
  • Proteomics
  • Genomics

Background:

  • High-throughput protein expression is crucial for structural genomics and proteomics.
  • Established methods include gene cloning (e.g., ligation-independent cloning) and recombinant expression in microbial hosts like Escherichia coli and Pichia pastoris.
  • These processes are increasingly automated and robotized.

Purpose of the Study:

  • To review optimized methods for high-throughput protein expression.
  • To highlight advancements in cell-free protein synthesis for large-scale production.
  • To discuss strategies for overcoming protein folding bottlenecks in structural studies.

Main Methods:

  • Utilizing robotic systems for gene cloning and recombinant protein expression.
  • Employing microbial hosts such as Escherichia coli and Pichia pastoris.
  • Leveraging cell-free protein synthesis systems for producing labeled proteins (e.g., for NMR and X-ray crystallography).
  • Implementing cell-based and cell-free screening methods to identify suitable protein samples for structure determination.

Main Results:

  • Gene cloning and recombinant expression in microbial systems are highly optimized and automated.
  • Cell-free systems enable large-scale protein production for specific applications like NMR and X-ray crystallography.
  • Protein folding remains a significant challenge in protein expression.
  • Screening methodologies have been developed to improve success rates in obtaining proteins for structural analysis.

Conclusions:

  • Optimized cloning and expression systems, including cell-free methods, facilitate high-throughput protein production.
  • Addressing protein folding challenges through effective screening is key to advancing structural genomics and proteomics.
  • Continued development in expression and screening technologies enhances the success rate of protein structure determination.