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From DNA to Protein

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...

Gene Evolution - Fast or Slow?

Gene Evolution - Fast or Slow?

The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
In contrast, regions which code...

Gene Evolution - Fast or Slow?

Gene Evolution - Fast or Slow?

The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
In contrast, regions which code...

Genome Size and the Evolution of New Genes

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While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.

Genome Size and the Evolution of New Genes

Genome Size and the Evolution of New Genes

While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.

The Central Dogma

The Central Dogma

The central dogma explains the flow of genetic information from DNA nucleotides to the amino acid sequence of proteins.
RNA is the Missing Link Between DNA and Proteins
In the early 1900s, scientists discovered that DNA stores all the information needed for cellular functions and that proteins perform most of these functions. However, the mechanisms of converting genetic information into functional proteins remained unknown for many years. Initially, it was believed that a single gene is...

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A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

A Facile Protocol to Generate Site-Specifically Acetylated Proteins in Escherichia Coli

Published on: December 9, 2017

Protein evolution with an expanded genetic code.

Chang C Liu¹, Antha V Mack, Meng-Lin Tsao

¹Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.

Proceedings of the National Academy of Sciences of the United States of America

|November 14, 2008

Summary

This summary is machine-generated.

Researchers developed a phage display system enabling directed evolution with unnatural amino acids. This synthetic genetic code allows selection of novel proteins, like antibodies, with enhanced functionality and binding properties.

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Area of Science:

Biotechnology
Molecular Biology
Protein Engineering

Background:

Traditional protein engineering relies on the 20 canonical amino acids.
Directed evolution aims to improve protein function but is limited by the natural amino acid repertoire.
Phage display is a powerful technique for selecting proteins with desired binding properties.

Purpose of the Study:

To develop a phage display system utilizing an expanded genetic code for directed evolution.
To enable the selection of proteins containing unnatural amino acids for improved function.
To demonstrate the utility of this system in generating high-affinity antibodies.

Main Methods:

Implementation of a phage display system incorporating an expanded genetic code.
Optimization of the system for various unnatural amino acids.
Randomization of specific antibody gene regions (V(H) CDR3) within a germline library.
Selection and characterization of phage-displayed antibodies against gp120.

Main Results:

The system successfully yielded proteins containing unnatural amino acids.
A phage-displayed antibody containing sulfotyrosine was selected.
This antibody demonstrated superior binding to gp120 compared to a known natural antibody.
The expanded genetic code conferred a selective advantage in directed evolution.

Conclusions:

An expanded "synthetic" genetic code can enhance directed evolution of proteins.
Unnatural amino acids can confer functional advantages, leading to improved protein properties.
This system offers a novel approach for engineering proteins with specific, enhanced functionalities.