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@ARTICLE{Tenhaef:890385,
author = {Tenhaef, Niklas and Stella, Robert and Frunzke, Julia and
Noack, Stephan},
title = {{A}utomated {R}ational {S}train {C}onstruction {B}ased on
{H}igh-{T}hroughput {C}onjugation},
journal = {ACS synthetic biology},
volume = {10},
number = {3},
issn = {2161-5063},
address = {Washington, DC},
publisher = {ACS},
reportid = {FZJ-2021-00918},
pages = {589–599},
year = {2021},
abstract = {Molecular cloning is the core of synthetic biology, as it
comprises the assembly of DNA and its expression in target
hosts. At present, however, cloning is most often a manual,
time-consuming, and repetitive process that highly benefits
from automation. The automation of a complete rational
cloning procedure, i.e., from DNA creation to expression in
the target host, involves the integration of different
operations and machines. Examples of such workflows are
sparse, especially when the design is rational (i.e., the
DNA sequence design is fixed and not based on randomized
libraries) and the target host is less genetically tractable
(e.g., not sensitive to heat-shock transformation). In this
study, an automated workflow for the rational construction
of plasmids and their subsequent conjugative transfer into
the biotechnological platform organism Corynebacterium
glutamicum is presented. The whole workflow is accompanied
by a custom-made software tool. As an application example, a
rationally designed library of transcription
factor-biosensors based on the regulator Lrp was constructed
and characterized. A sensor with an improved dynamic range
was obtained, and insights from the screening provided
evidence for a dual regulator function of C. glutamicum
Lrp.},
cin = {IBG-1},
ddc = {570},
cid = {I:(DE-Juel1)IBG-1-20101118},
pnm = {2171 - Biological and environmental resources for
sustainable use (POF4-217)},
pid = {G:(DE-HGF)POF4-2171},
typ = {PUB:(DE-HGF)16},
pubmed = {33593066},
UT = {WOS:000631444600015},
doi = {10.1021/acssynbio.0c00599},
url = {https://juser.fz-juelich.de/record/890385},
}