scholarly journals UGA Read-Through Artifacts—When Popular Gene Expression Systems Need a pATCH

BioTechniques ◽  
1998 ◽  
Vol 24 (5) ◽  
pp. 789-794 ◽  
Author(s):  
Gavin MacBeath ◽  
Peter Kast
Keyword(s):  
Gene Expression ◽  
Expression Systems

scholarly journals On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

2021 ◽  
Author(s):  
Philipp Moritz Fricke ◽  
Angelika Klemm ◽  
Michael Bott ◽  
Tino Polen
Keyword(s):  
Gene Expression ◽  
Acetic Acid ◽  
Literature Search ◽  
Target Gene ◽  
Target Genes ◽  
Recombinant Dna ◽  
Acetic Acid Bacteria ◽  
Homoserine Lactone ◽  
Expression Systems ◽  
Target Gene Expression

Abstract Acetic acid bacteria (AAB) are valuable biocatalysts for which there is growing interest in understanding their basics including physiology and biochemistry. This is accompanied by growing demands for metabolic engineering of AAB to take advantage of their properties and to improve their biomanufacturing efficiencies. Controlled expression of target genes is key to fundamental and applied microbiological research. In order to get an overview of expression systems and their applications in AAB, we carried out a comprehensive literature search using the Web of Science Core Collection database. The Acetobacteraceae family currently comprises 49 genera. We found overall 6097 publications related to one or more AAB genera since 1973, when the first successful recombinant DNA experiments in Escherichia coli have been published. The use of plasmids in AAB began in 1985 and till today was reported for only nine out of the 49 AAB genera currently described. We found at least five major expression plasmid lineages and a multitude of further expression plasmids, almost all enabling only constitutive target gene expression. Only recently, two regulatable expression systems became available for AAB, an N-acyl homoserine lactone (AHL)-inducible system for Komagataeibacter rhaeticus and an l-arabinose-inducible system for Gluconobacter oxydans. Thus, after 35 years of constitutive target gene expression in AAB, we now have the first regulatable expression systems for AAB in hand and further regulatable expression systems for AAB can be expected. Key points • Literature search revealed developments and usage of expression systems in AAB. • Only recently 2 regulatable plasmid systems became available for only 2 AAB genera. • Further regulatable expression systems for AAB are in sight.

Construction of gene expression systems in insect cell lines using promoters from the silkworm, Bombyx mori

2008 ◽  
Vol 133 (1) ◽  
pp. 9-17 ◽  
Author(s):  
Jae Man Lee ◽  
Masateru Takahashi ◽  
Hiroaki Mon ◽  
Hitoshi Mitsunobu ◽  
Katsumi Koga ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bombyx Mori ◽  
Cell Lines ◽  
Insect Cell ◽  
Expression Systems ◽  
Insect Cell Lines ◽  
Silkworm Bombyx Mori

Cytotoxic effect of doxycycline and its implications for tet-on gene expression systems

2003 ◽  
Vol 318 (1) ◽  
pp. 152-154 ◽  
Author(s):  
Gennady Ermak ◽  
Vincenzo J Cancasci ◽  
Kelvin J.A Davies
Keyword(s):  
Gene Expression ◽  
Cytotoxic Effect ◽  
Expression Systems

scholarly journals An overview on current molecular tools for heterologous gene expression in Trichoderma

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Irene Tomico-Cuenca ◽  
Robert L. Mach ◽  
Astrid R. Mach-Aigner ◽  
Christian Derntl
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
Protein Secretion ◽  
Heterologous Gene Expression ◽  
Heterologous Gene ◽  
Molecular Tools ◽  
Expression Systems ◽  
Industrial Enzymes ◽  
Nutrient Sources ◽  
Transformation Methods

AbstractFungi of the genus Trichoderma are routinely used as biocontrol agents and for the production of industrial enzymes. Trichoderma spp. are interesting hosts for heterologous gene expression because their saprotrophic and mycoparasitic lifestyles enable them to thrive on a large number of nutrient sources and some members of this genus are generally recognized as safe (GRAS status). In this review, we summarize and discuss several aspects involved in heterologous gene expression in Trichoderma, including transformation methods, genome editing strategies, native and synthetic expression systems and implications of protein secretion. This review focuses on the industrial workhorse Trichoderma reesei because this fungus is the best-studied member of this genus for protein expression and secretion. However, the discussed strategies and tools can be expected to be transferable to other Trichoderma species.

scholarly journals Plasmids for Independently Tunable, Low-Noise Expression of Two Genes

mSphere ◽  
2019 ◽  
Vol 4 (3) ◽  
Author(s):  
João P. N. Silva ◽  
Soraia Vidigal Lopes ◽  
Diogo J. Grilo ◽  
Zach Hensel
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
Low Noise ◽  
Expression Systems ◽  
Extrinsic Noise ◽  
Gene Expression Noise ◽  
Expression Noise ◽  
Foreign Proteins ◽  
Noise Limit ◽  
Cell Variation

ABSTRACTSome microbiology experiments and biotechnology applications can be improved if it is possible to tune the expression of two different genes at the same time with cell-to-cell variation at or below the level of genes constitutively expressed from the chromosome (the “extrinsic noise limit”). This was recently achieved for a single gene by exploiting negative autoregulation by the tetracycline repressor (TetR) and bicistronic gene expression to reduce gene expression noise. We report new plasmids that use the same principles to achieve simultaneous, low-noise expression for two genes inEscherichia coli. The TetR system was moved to a compatible plasmid backbone, and a system based on thelacrepressor (LacI) was found to also exhibit gene expression noise below the extrinsic noise limit. We characterized gene expression mean and noise across the range of induction levels for these plasmids, applied the LacI system to tune expression for single-molecule mRNA detection under two different growth conditions, and showed that two plasmids can be cotransformed to independently tune expression of two different genes.IMPORTANCEMicrobiologists often express foreign proteins in bacteria in order study them or to use bacteria as a microbial factory. Usually, this requires controlling the number of foreign proteins expressed in each cell, but for many common protein expression systems, it is difficult to “tune” protein expression without large cell-to-cell variation in expression levels (called “noise” in protein expression). This work describes two protein expression systems that can be combined in the same cell, with tunable expression levels and very low protein expression noise. One new system was used to detect single mRNA molecules by fluorescence microscopy, and the two systems were shown to be independent of each other. These protein expression systems may be useful in any experiment or biotechnology application that can be improved with low protein expression noise.

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