A split intein T7 RNA polymerase for transcriptional AND-logic.
Details
Download: Pub_NAR2014.pdf (219.96 [Ko])
State: Public
Version: Final published version
State: Public
Version: Final published version
Serval ID
serval:BIB_BAF1E83CDB91
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
A split intein T7 RNA polymerase for transcriptional AND-logic.
Journal
Nucleic Acids Research
ISSN
1362-4962 (Electronic)
ISSN-L
0305-1048
Publication state
Published
Issued date
2014
Volume
42
Number
19
Pages
12322-12328
Language
english
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.
Keywords
DNA-Directed RNA Polymerases/chemistry, DNA-Directed RNA Polymerases/metabolism, Escherichia coli/genetics, Inteins, Molecular Sequence Data, Trans-Splicing, Transcription, Genetic, Viral Proteins/chemistry, Viral Proteins/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
01/02/2017 21:21
Last modification date
20/08/2019 15:28