Schaerli, YolandaGili, MaguiIsalan, Mark2025-01-132025-01-132014Schaerli Y, Gili M, Isalan M. A split intein T7 RNA polymerase for transcriptional AND-logic. Nucleic Acids Res. 2014 Oct 29;42(19):12322-8. DOI: 10.1093/nar/gku8840305-1048http://hdl.handle.net/10230/69064Synthetic 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.application/pdfeng© 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.Reacció en cadena de la polimerasaProcariotesProteïnesinfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1093/nar/gku884info:eu-repo/semantics/openAccess