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Research

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.

Publications

Citation

BibTex format

@article{Aw:2016:10.1186/s12934-016-0432-8,
author = {Aw, R and Polizzi, KM},
doi = {10.1186/s12934-016-0432-8},
journal = {Microbial Cell Factories},
title = {Liquid PTVA: A faster and cheaper alternative for generating multi-copy clones in Pichia pastoris},
url = {http://dx.doi.org/10.1186/s12934-016-0432-8},
volume = {15},
year = {2016}
}

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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - BACKGROUND:Multiple cognate gene copy clones have often been used in order to increase the yield of recombinant protein expression in the yeast Pichia pastoris. The method of posttransformational vector amplification (PTVA) has allowed for the efficient generation of multi-copy clones in P. pastoris. However, despite its relative ease and success, this process can be expensive and time consuming.RESULTS:We have developed a modified version of PTVA, called Liquid PTVA, which allows for faster and cheaper selection of multi-copy clones. Cultures are grown in liquid medium with only a final selection carried out on agar plates, reducing overall antibiotic usage and increasing the speed of clone amplification. In addition, it was established that starting PTVA with a single copy clone resulted in higher copy number strains for both traditional plate PTVA and liquid PTVA. Furthermore, using the Zeocin selection marker in liquid PTVA results in strains with higher growth rates, which could be beneficial for recombinant protein production processes.CONCLUSIONS:We present a methodology for creating multi-copy clones that can be achieved over 12 days instead of the traditional 45 and at approximately half the cost.
AU  - Aw,R
AU  - Polizzi,KM
DO  - 10.1186/s12934-016-0432-8
PY  - 2016///
SN  - 1475-2859
TI  - Liquid PTVA: A faster and cheaper alternative for generating multi-copy clones in Pichia pastoris
T2  - Microbial Cell Factories
UR  - http://dx.doi.org/10.1186/s12934-016-0432-8
UR  - http://hdl.handle.net/10044/1/29028
VL  - 15
ER  -

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