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Synthetic Biology underpins advances in the bioeconomy
Biological systems - including the simplest cells - exhibit a broad range of functions to thrive in their environment. Research in the Imperial College Centre for Synthetic Biology is focused on the possibility of engineering the underlying biochemical processes to solve many of the challenges facing society, from healthcare to sustainable energy. In particular, we model, analyse, design and build biological and biochemical systems in living cells and/or in cell extracts, both exploring and enhancing the engineering potential of biology.
As part of our research we develop novel methods to accelerate the celebrated Design-Build-Test-Learn synthetic biology cycle. As such research in the Centre for Synthetic Biology highly multi- and interdisciplinary covering computational modelling and machine learning approaches; automated platform development and genetic circuit engineering ; multi-cellular and multi-organismal interactions, including gene drive and genome engineering; metabolic engineering; in vitro/cell-free synthetic biology; engineered phages and directed evolution; and biomimetics, biomaterials and biological engineering.
@article{Aw:2020:10.1002/cpps.115,
author = {Aw, R and Spice, AJ and Polizzi, K},
doi = {10.1002/cpps.115},
journal = {Current protocols in protein science},
title = {Methods for expression of recombinant proteins using a Pichia pastoris cell-free system},
url = {http://dx.doi.org/10.1002/cpps.115},
volume = {102},
year = {2020}
}
TY - JOUR
AB - Cellfree protein synthesis is a powerful tool for engineering biology and has been utilized in many diverse applications, from biosensing and protein prototyping to biomanufacturing and the design of metabolic pathways. By exploiting host cellular machinery decoupled from cellular growth, proteins can be produced in vitro both on demand and rapidly. Eukaryotic cellfree platforms are often neglected due to perceived complexity and low yields relative to their prokaryotic counterparts, despite providing a number of advantageous properties. The yeast Pichia pastoris (also known as Komagataella phaffii) is a particularly attractive eukaryotic host from which to generate cellfree extracts, due to its ability to grow to high cell densities with high volumetric productivity, genetic tractability for strain engineering, and ability to perform posttranslational modifications. Here, we describe methods for conducting cellfree protein synthesis using P. pastoris as the host, from preparing the cell lysates to protocols for both coupled and linked transcriptiontranslation reactions. By providing these methodologies, we hope to encourage the adoption of the platform by new and experienced users alike.
AU - Aw,R
AU - Spice,AJ
AU - Polizzi,K
DO - 10.1002/cpps.115
PY - 2020///
SN - 1934-3655
TI - Methods for expression of recombinant proteins using a Pichia pastoris cell-free system
T2 - Current protocols in protein science
UR - http://dx.doi.org/10.1002/cpps.115
UR - http://hdl.handle.net/10044/1/83872
VL - 102
ER -