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Related Concept Videos

Ribozymes02:47

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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
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Within a biological system, the DNA encodes the RNA, and the nucleotide sequence in the RNA further defines the amino acid sequence in the protein. This is referred to as “The Central Dogma of Molecular Biology” - a term coined by Francis Crick.  Central dogma is a firm principle in biology that defines the flow of genetic information within any life form. The two fundamental steps in central dogma are - transcription and translation.
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The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
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Related Experiment Video

Updated: Apr 21, 2026

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
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RApid Parallel Protein EvaluatoR (RAPPER), from gene to enzyme function in one day.

L T Quertinmont1, R Orru, S Lutz

  • 1Department of Chemistry, Emory University, 1515 Dickey Drive, Atlanta, GA 30322, USA. sal2@emory.edu.

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Summary

Cell-free systems enable rapid protein variant analysis. This study introduces a DNA template method for evaluating genetic variations

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Synthetic Biology

Background:

  • Cell-free transcription-translation systems provide a flexible platform for studying protein function.
  • Genetic variations can significantly alter protein properties, necessitating efficient evaluation methods.

Purpose of the Study:

  • To develop a protocol for preparing linear DNA templates for cell-free systems.
  • To enable fast and semi-quantitative assessment of amino acid variations in proteins.
  • To evaluate catalytic activity and stereo-selectivity of Old Yellow Enzyme variants.

Main Methods:

  • Development of a protocol for linear, mutagenic DNA template preparation.
  • Utilizing the PURE system (Protein synthesis Using Recombinant Elements) for cell-free expression.
  • Semi-quantitative evaluation of enzyme variants' catalytic activity and stereo-selectivity.

Main Results:

  • A protocol for preparing linear DNA templates compatible with the PURE system was established.
  • The method allows for rapid assessment of amino acid variations.
  • The impact of variations on Old Yellow Enzyme's catalytic activity and stereo-selectivity was evaluated.

Conclusions:

  • The developed protocol offers an efficient approach for analyzing genetic variations' effects on protein function.
  • This method facilitates the study of both native and engineered enzyme variants.
  • Cell-free systems coupled with optimized DNA templates are powerful tools for protein engineering and functional studies.