A split intein T7 RNA polymerase for transcriptional AND-logic. Issue 19 (27th September 2014)

• Record Type: Journal Article
• Authors: Schaerli, Yolanda; Gili, Magüi; Isalan, Mark
• Is Part Of: Nucleic acids research. Volume 42:Issue 19(2014)
• Page Start: 12322
• Page End: 12328
• Publication Date: 2014-09-27
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gku884
• Languages: English
• ISSNs: 0305-1048

Abstract

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.