TY - JOUR
T1 - A positive feedback-based gene circuit to increase the production of a membrane protein
AU - Bansal, Karan
AU - Yang, Ke
AU - Nistala, Goutam J.
AU - Gennis, Robert B.
AU - Bhalerao, Kaustubh D.
PY - 2010/5/25
Y1 - 2010/5/25
N2 - Background: Membrane proteins are an important class of proteins, playing a key role in many biological processes, and are a promising target in pharmaceutical development. However, membrane proteins are often difficult to produce in large quantities for the purpose of crystallographic or biochemical analyses.Results: In this paper, we demonstrate that synthetic gene circuits designed specifically to overexpress certain genes can be applied to manipulate the expression kinetics of a model membrane protein, cytochrome bd quinol oxidase in E. coli, resulting in increased expression rates. The synthetic circuit involved is an engineered, autoinducer-independent variant of the lux operon activator LuxR from V. fischeri in an autoregulatory, positive feedback configuration.Conclusions: Our proof-of-concept experiments indicate a statistically significant increase in the rate of production of the bd oxidase membrane protein. Synthetic gene networks provide a feasible solution for the problem of membrane protein production.
AB - Background: Membrane proteins are an important class of proteins, playing a key role in many biological processes, and are a promising target in pharmaceutical development. However, membrane proteins are often difficult to produce in large quantities for the purpose of crystallographic or biochemical analyses.Results: In this paper, we demonstrate that synthetic gene circuits designed specifically to overexpress certain genes can be applied to manipulate the expression kinetics of a model membrane protein, cytochrome bd quinol oxidase in E. coli, resulting in increased expression rates. The synthetic circuit involved is an engineered, autoinducer-independent variant of the lux operon activator LuxR from V. fischeri in an autoregulatory, positive feedback configuration.Conclusions: Our proof-of-concept experiments indicate a statistically significant increase in the rate of production of the bd oxidase membrane protein. Synthetic gene networks provide a feasible solution for the problem of membrane protein production.
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U2 - 10.1186/1754-1611-4-6
DO - 10.1186/1754-1611-4-6
M3 - Article
C2 - 20500847
AN - SCOPUS:77952491659
SN - 1754-1611
VL - 4
JO - Journal of Biological Engineering
JF - Journal of Biological Engineering
M1 - 6
ER -