Phage Assisted Continuous Evolution (PACE)
Phage-assisted continuous evolution (PACE) is a system that enables the continuous directed evolution of gene-encoded molecules that can be linked to protein production in Escherichia coli. During PACE, evolving genes are transferred from host cell to host cell through a modified bacteriophage life cycle in a manner that is dependent on the activity of interest.
According to the experimental protocol provided by David Liu (Fig 1), the laboratory of Profacgen optimizes the experimental scheme to provide different guiding strategies for PACE screening for different types of polymerase I, and can now provide advanced and personalized PACE service for scientific research institutions or companies in need.
Fig 1 Overview of the the PACE system [1].
Currently, Profacgen has successfully used directed evolution to identify protein variants resistant to high temperature, denaturants or proteases, screen protein libraries in whole cells or cell lysates, identify highly active variants, and produce proteins with excellent expression.
Profacgen also improved the phage-assisted continuous directed evolution system and method, which included phages carrying target genes to be evolved, multiple helper plasmids supporting the proliferation of different phages before and after evolution, and plasmids inducing mutations. The placement of different Hp in different host bacteria can effectively improve the efficiency of phage proliferation and evolution, while avoiding the mutual interference between different genetic elements, which has good practicability and can be used for the directional evolution of a variety of genes.
Workflow of PACE service in Profacgen:
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Reference:
[1] Esvelt, Kevin M.; Carlson, Jacob C.; Liu, David R. (2011). A system for the continuous directed evolution of biomolecules. , 472(7344), 499–503. doi:10.1038/nature09929