Article: article from journal or magazin.
A split intein T7 RNA polymerase for transcriptional AND-logic.
Nucleic Acids Research
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.
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
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