caa a22 4500 605501467 CHVBK 20210128100601.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6610-5 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6610-5 Development of a regulatable plasmid-based gene expression system for Clostridium thermocellum [Elektronische Daten] [Elizabeth Mearls, Daniel Olson, Christopher Herring, Lee Lynd] Clostridium thermocellum can rapidly solubilize cellulose and produces ethanol as an end product of its metabolism. As such, it is a candidate for bioethanol production from plant matter. In this study, we developed an inducible expression system for C. thermocellum based on its native celC operon. We enhanced expression over the native operon structure by placing the repressor gene, glyR3, immediately after the celC promoter, and expressing the target gene after glyR3. Upon the addition of the inducer substrate, laminaribiose, an approximately 40-fold increase in gene expression was obtained using the test gene spo0A. Furthermore, induction of the sporulation histidine kinase, clo1313_1942, increased sporulation frequency by approximately 10,000-fold relative to an uninduced control. We have also shown that the laminaribiose (β1-3-linked carbon source) utilization pathway is not catabolite repressed by cellobiose, a β1-4-linked carbon source frequently used for C. thermocellum cultivation in laboratory conditions. Selective expression of target genes has the potential to inform metabolic engineering strategies as well as increase fundamental understanding of C. thermocellum biology. Springer-Verlag Berlin Heidelberg (outside the USA), 2015 Laminaribiose nationallicence Inducible promoter nationallicence Biofuels nationallicence Spore formation nationallicence Mearls Elizabeth Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut Olson Daniel Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut Herring Christopher Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut Lynd Lee Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7589-7599 0175-7598 99:18<7589 2015 99 253 https://doi.org/10.1007/s00253-015-6610-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6610-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mearls Elizabeth Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut NATIONALLICENCE 700 1- Olson Daniel Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut NATIONALLICENCE 700 1- Herring Christopher Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut NATIONALLICENCE 700 1- Lynd Lee Thayer School of Engineering, Dartmouth College, 03755, Hanover, NH, USA aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7589-7599 0175-7598 99:18<7589 2015 99 253