A split intein T7 RNA polymerase for transcriptional AND-logic
File(s)43 T7 split intein nar.gku884.full.pdf (227.11 KB)
Published version
Author(s)
Schaerli, Y
Gili, M
Isalan, M
Type
Journal Article
Abstract
Synthetic biology has developed numerous parts for building synthetic gene circuits. However, few parts have been described for prokaryotes to integrate two signals at a promoter in an AND fashion, i.e. the promoter is only activated in the presence of both signals. Here we present a new part for this function: a split intein T7 RNA polymerase. We divide T7 RNA polymerase into two expression domains and fuse each to a split intein. Only when both domains are expressed does the split intein mediate protein trans-splicing, yielding a full-length T7 RNA polymerase that can transcribe genes via a T7 promoter. We demonstrate an AND gate with the new part: the signal-to-background ratio is very high, resulting in an almost digital signal. This has utility for more complex circuits and so we construct a band-pass filter in Escherichia coli. The split intein approach should be widely applicable for engineering artificial gene circuit parts.
Date Issued
2014-09-27
Citation
Nucleic Acids Research, 2014
ISSN
1362-4962
Publisher
Oxford University Press (OUP)
Journal / Book Title
Nucleic Acids Research
Copyright Statement
© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
License URL
Description
03.11.14 KB. Ok to add published version to spiral, OA paper under cc license
Identifier
http://nar.oxfordjournals.org/cgi/content/full/gku884?%20ijkey=pakHfX8umQZOeVk&keytype=ref