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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

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 helps...
Leaky Scanning02:28

Leaky Scanning

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 stands for...
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

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 addition of a...
Translational Regulation01:29

Translational Regulation

Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
What is Gene Expression?01:36

What is Gene Expression?

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 processed and...
From DNA to Protein03:06

From DNA to Protein

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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Updated: Jun 7, 2026

Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling
06:58

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Published on: October 7, 2021

Tuning protein expression using synonymous codon libraries targeted to the 5' mRNA coding region.

Lise Goltermann1, Martin Borch Jensen, Thomas Bentin

  • 1Department of Cellular and Molecular Medicine, The Panum Institute, University of Copenhagen, Blegdamsvej 3b, 2200 Copenhagen, Denmark.

Protein Engineering, Design & Selection : PEDS
|November 5, 2010
PubMed
Summary

Synthetic sequences in the 5' mRNA coding region of bacteria can control protein expression levels. This method tunes recombinant protein production over 300-fold without altering amino acid sequences, impacting gene expression research.

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Last Updated: Jun 7, 2026

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Cell Based Assays of SINEUP Non-coding RNAs That Can Specifically Enhance mRNA Translation
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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
11:51

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

Area of Science:

  • Molecular Biology
  • Bacterial Genetics
  • Protein Expression

Background:

  • The 5' mRNA coding region influences translation efficiency in bacteria.
  • Understanding regulatory elements in this region is crucial for controlling protein synthesis.

Purpose of the Study:

  • To investigate the impact of synthetic sequences in the 5' mRNA coding region on recombinant protein expression in Escherichia coli.
  • To develop a method for tuning protein expression levels by modifying the 5' coding sequence.

Main Methods:

  • Introduction of synthetic sequences into the 5' mRNA coding region of a reporter gene.
  • Utilizing a synonymous codon library to modify the 5' coding sequence while preserving amino acid identity.
  • Measuring green fluorescent protein (GFP) expression levels to assess translation output.

Main Results:

  • Synthetic sequences dramatically altered GFP expression, ranging from near abolition to enhancement.
  • Protein stability was not the primary factor for expression differences, suggesting RNA-level regulation.
  • A synonymous codon library enabled a ~300-fold tuning range of protein expression.

Conclusions:

  • Features within the 5' mRNA coding region, near the start codon, are critical determinants of translation output in bacteria.
  • This RNA-level regulatory approach offers a simple and broadly applicable tool for controlling recombinant and endogenous gene expression.