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Synthetic tunable amplifying buffer circuit in E. coli.

Abstract：

:While predictable design of a genetic circuit's output is a major goal of synthetic biology, it remains a significant challenge because DNA binding sites in the cell affect the concentration of available transcription factors (TF). To mitigate this problem, we propose to use a TF that results from the (reversible) phosphorylation of protein substrate as a circuit's output. We demonstrate that by comparatively increasing the amounts of substrate and phosphatase, the TF concentration becomes robust to the presence of DNA binding sites and can be kept at a desired value. The circuit's input/output gain can, in turn, be tuned by changing the relative amounts of the substrate and phosphatase, realizing an amplifying buffer circuit with tunable gain. In our experiments in E. coli, we employ phospho-NRI as the output TF, phosphorylated by the NRII kinase, and dephosphorylated by the NRII phosphatase. Amplifying buffer circuits such as ours could be used to insulate a circuit's output from the context, bringing synthetic biology one step closer to modular design.

journal_name

ACS Synth Biol

journal_title

ACS synthetic biology

authors

Nilgiriwala KS,Jiménez J,Rivera PM,Del Vecchio D

doi

10.1021/sb5002533

subject

Has Abstract

pub_date

2015-05-15 00:00:00

pages

577-84

issue

5

issn

2161-5063

journal_volume

4

pub_type

杂志文章

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