A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
GreA and GreB Enhance Expression of Escherichia coli RNA Polymerase Promoters in a Reconstituted Transcription–Translation System




Julkaisun tekijät: Lea L. de Maddalena, Henrike Niederholtmeyer, Matti Turtola, Zoe N. Swank, Georgiy A. Belogurov, and Sebastian J. Maerkl
Kustantaja: American Chemical Society
Julkaisuvuosi: 2016
Journal: ACS Synthetic Biology
Tietokannassa oleva lehden nimi: ACS SYNTHETIC BIOLOGY
Lehden akronyymi: ACS SYNTH BIOL
Volyymi: 5
Julkaisunumero: 9
eISSN: 2161-5063

Tiivistelmä

Cell-free environments are becoming viable alternatives for implementing
biological networks in synthetic biology. The reconstituted cell-free
expression system (PURE) allows characterization of genetic networks
under defined conditions but its applicability to native bacterial
promoters and endogenous genetic networks is limited due to the poor
transcription rate of Escherichia coli RNA polymerase in this
minimal system. We found that addition of transcription elongation
factors GreA and GreB to the PURE system increased transcription rates
of E. coli RNA polymerase from sigma factor 70 promoters up to
6-fold and enhanced the performance of a genetic network. Furthermore,
we reconstituted activation of natural E. coli promoters
controlling flagella biosynthesis by the transcriptional activator FlhDC
and sigma factor 28. Addition of GreA/GreB to the PURE system allows
efficient expression from natural and synthetic E. coli promoters
and characterization of their regulation in minimal and defined
reaction conditions, making the PURE system more broadly applicable to
study genetic networks and bottom-up synthetic biology.



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Last updated on 2019-29-01 at 16:11