109Synthetic Biology: Parts, Devices and Applications, First Edition. Edited by Christina Smolke.
© 2018 Wiley-VCH Verlag GmbH & Co. KGaA. Published 2018 by Wiley-VCH Verlag GmbH & Co. KGaA.
6
The implementation of synthetic genetic circuits requires precise control of the
expression of every gene involved. This can be achieved by choosing promoters
that appropriately modulate transcription initiation in terms of intensity and
duration in response to specific stimuli. In nature, promoters couple gene
expression to the internal status of the cell and to the external conditions of the
environment. Here, we describe Saccharomyces cerevisiae promoters. The char-
acterization of the structural and functional features of natural promoters has
been crucial for their application. Moreover, this knowledge led to the imple-
mentation of synthetic promoters displaying novel regulatory properties.
6.1 Introduction
The characterization of Saccharomyces cerevisiae promoters began by using
them to drive expression of reporter genes. Systematic truncations and deletions
of the promoter region of these constructs revealed that yeast promoters share a
common modular structure [1, 2]. Each module has a defined role in the stimula-
tion and regulation of transcription initiation [3, 4]. The characterization of
natural promoters allowed their use in controlling the expression of heterolo-
gous genes [5–7].
Today, a large selection of well‐characterized natural promoters is routinely
exploited for controlling transcription in yeast [8, 9]. Although these promoters
span a wide range of transcription initiation efficiencies, they usually do not
cover them homogeneously; that is, most promoters display either very weak or
very strong activity. Moreover, natural yeast promoters cannot be used to
build orthogonal systems, since they are intimately linked to metabolism. These
two limitations are overcome by constructing synthetic promoters, whose
strength can be finely tuned and whose regulation can be independent of
the metabolism.
Constitutive and Regulated Promoters in Yeast: How
to Design and Make Use of Promoters in S. cerevisiae
Diana S. M. Ottoz1,2 and Fabian Rudolf^2
(^1) ETH Zurich, Department of Biosystems Science and Engineering, Mattenstrasse 26, 4058 Basel, Switzerland
(^2) Yale University, Department of Molecular Biophysics and Biochemistry, 333 Cedar street SHM C-111, New Haven,
CT, 06520, USA