Jasmine De Baets, Irene Parmentier, Brecht De Paepe, Marjan De Mey
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引用次数: 0
Abstract
The interesting features of quorum sensing systems make them very appealing for synthetic biology applications. The first steps have already been taken in characterizing these systems to accelerate their successful implementation. In this work, we explore the next step, tuning. Multiple strategies exist for tuning genetic circuits and quorum sensing systems in general, where the most straightforward solution is to vary the expression level of the synthase or transcription factor. However, these tuning possibilities can be broadened by expanding the variety of quorum sensing-regulated promoters. Here, we expand the range of tuning possibilities by constructing promoter libraries for PlasI (LasI/LasR system) and PesaR/PesaS (EsaI/EsaR system), enabling direct modulation of downstream gene expression levels. Next, the quorum sensing synthetic biology toolbox was further expanded with the creation of a new-to-nature genetic part, the hybrid promoter, which requires two autoinducers for activation, namely 3-oxo-hexanoyl- and 3-oxo-dodecanoyl-homoserine lactone. This promoter was optimized and applied to design a multistage genetic switch capable of creating a growth stage followed by two sequential production stages. Additionally, the circuit was optimized by incorporating the new promoter variants created in this study. This complex genetic circuit demonstrates the true potential of these quorum sensing systems for achieving various cell-density related dynamic circuits.
期刊介绍:
New Biotechnology is the official journal of the European Federation of Biotechnology (EFB) and is published bimonthly. It covers both the science of biotechnology and its surrounding political, business and financial milieu. The journal publishes peer-reviewed basic research papers, authoritative reviews, feature articles and opinions in all areas of biotechnology. It reflects the full diversity of current biotechnology science, particularly those advances in research and practice that open opportunities for exploitation of knowledge, commercially or otherwise, together with news, discussion and comment on broader issues of general interest and concern. The outlook is fully international.
The scope of the journal includes the research, industrial and commercial aspects of biotechnology, in areas such as: Healthcare and Pharmaceuticals; Food and Agriculture; Biofuels; Genetic Engineering and Molecular Biology; Genomics and Synthetic Biology; Nanotechnology; Environment and Biodiversity; Biocatalysis; Bioremediation; Process engineering.