High throughput construction and small scale expression screening of multi-tag vectors in Escherichia coli

Methods ◽  
2011 ◽  
Vol 55 (1) ◽  
pp. 29-37 ◽  
Author(s):  
Louise E. Bird
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
Small Scale ◽  
Expression Screening

High-throughput protein expression screening and purification in Escherichia coli

Methods ◽  
2011 ◽  
Vol 55 (1) ◽  
pp. 65-72 ◽  
Author(s):  
Renaud Vincentelli ◽  
Agnès Cimino ◽  
Arie Geerlof ◽  
Atsushi Kubo ◽  
Yutaka Satou ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Expression ◽  
High Throughput ◽  
Expression Screening

scholarly journals Molecules incorporating a benzothiazole core scaffold inhibit the N-myristoyltransferase of Plasmodium falciparum

2007 ◽  
Vol 408 (2) ◽  
pp. 173-180 ◽  
Author(s):  
Paul W. Bowyer ◽  
Ruwani S. Gunaratne ◽  
Munira Grainger ◽  
Chrislaine Withers-Martinez ◽  
Sasala R. Wickramsinghe ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Plasmodium Falciparum ◽  
High Throughput ◽  
High Throughput Screening ◽  
Small Scale ◽  
Expression And Purification ◽  
Purification System ◽  
Pure Protein ◽  
Ic50 Values

Recombinant N-myristoyltransferase of Plasmodium falciparum (termed PfNMT) has been used in the development of a SPA (scintillation proximity assay) suitable for automation and high-throughput screening of inhibitors against this enzyme. The ability to use the SPA has been facilitated by development of an expression and purification system which yields considerably improved quantities of soluble active recombinant PfNMT compared with previous studies. Specifically, yields of pure protein have been increased from 12 μg·l−1 to >400 μg·l−1 by use of a synthetic gene with codon usage optimized for expression in an Escherichia coli host. Preliminary small-scale ‘piggyback’ inhibitor studies using the SPA have identified a family of related molecules containing a core benzothiazole scaffold with IC50 values <50 μM, which demonstrate selectivity over human NMT1. Two of these compounds, when tested against cultured parasites in vitro, reduced parasitaemia by >80% at a concentration of 10 μM.

scholarly journals Fully Automated Cultivation of Adipose-Derived Stem Cells in the StemCellDiscovery—A Robotic Laboratory for Small-Scale, High-Throughput Cell Production Including Deep Learning-Based Confluence Estimation

Processes ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 575
Author(s):  
Jelena Ochs ◽  
Ferdinand Biermann ◽  
Tobias Piotrowski ◽  
Frederik Erkens ◽  
Bastian Nießing ◽  
...  
Keyword(s):  
Stem Cells ◽  
Deep Learning ◽  
High Throughput ◽  
High Speed ◽  
New Technologies ◽  
Human Mesenchymal Stem Cells ◽  
Simulation Software ◽  
System Capacity ◽  
Small Scale ◽  
Production Environment

Laboratory automation is a key driver in biotechnology and an enabler for powerful new technologies and applications. In particular, in the field of personalized therapies, automation in research and production is a prerequisite for achieving cost efficiency and broad availability of tailored treatments. For this reason, we present the StemCellDiscovery, a fully automated robotic laboratory for the cultivation of human mesenchymal stem cells (hMSCs) in small scale and in parallel. While the system can handle different kinds of adherent cells, here, we focus on the cultivation of adipose-derived hMSCs. The StemCellDiscovery provides an in-line visual quality control for automated confluence estimation, which is realized by combining high-speed microscopy with deep learning-based image processing. We demonstrate the feasibility of the algorithm to detect hMSCs in culture at different densities and calculate confluences based on the resulting image. Furthermore, we show that the StemCellDiscovery is capable of expanding adipose-derived hMSCs in a fully automated manner using the confluence estimation algorithm. In order to estimate the system capacity under high-throughput conditions, we modeled the production environment in a simulation software. The simulations of the production process indicate that the robotic laboratory is capable of handling more than 95 cell culture plates per day.

scholarly journals Quantification of enterohemorrhagic Escherichia coli O157:H7 protein abundance by high-throughput proteome

PLoS ONE ◽  
2018 ◽  
Vol 13 (12) ◽  
pp. e0208520 ◽  
Author(s):  
Wanderson Marques Da Silva ◽  
Jinlong Bei ◽  
Natalia Amigo ◽  
María Pía Valacco ◽  
Ariel Amadio ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
Enterohemorrhagic Escherichia Coli ◽  
Escherichia Coli O157 ◽  
Protein Abundance

Co-expressional conservation in virulence and stress related genes of three Gammaproteobacterial species: Escherichia coli, Salmonella enterica and Pseudomonas aeruginosa

2015 ◽  
Vol 11 (11) ◽  
pp. 3137-3148
Author(s):  
Nazanin Hosseinkhan ◽  
Peyman Zarrineh ◽  
Hassan Rokni-Zadeh ◽  
Mohammad Reza Ashouri ◽  
Ali Masoudi-Nejad
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Systems Biology ◽  
Gene Expression Data ◽  
High Throughput ◽  
Expression Data ◽  
P Gene ◽  
Stress Related Genes ◽  
High Throughput Gene Expression

Gene co-expression analysis is one of the main aspects of systems biology that uses high-throughput gene expression data.

scholarly journals A High-Throughput Approach To Identify Compounds That Impair Envelope Integrity in Escherichia coli

2016 ◽  
Vol 60 (10) ◽  
pp. 5995-6002 ◽  
Author(s):  
Kristin R. Baker ◽  
Bimal Jana ◽  
Henrik Franzyk ◽  
Luca Guardabassi
Keyword(s):  
Escherichia Coli ◽  
High Throughput ◽  
High Throughput Screening ◽  
Gram Negative Bacteria ◽  
Chromogenic Substrate ◽  
Intrinsic Resistance ◽  
Gram Negative ◽  
Content Type ◽  
E Coli ◽  
Screening Platform

ABSTRACTThe envelope of Gram-negative bacteria constitutes an impenetrable barrier to numerous classes of antimicrobials. This intrinsic resistance, coupled with acquired multidrug resistance, has drastically limited the treatment options against Gram-negative pathogens. The aim of the present study was to develop and validate an assay for identifying compounds that increase envelope permeability, thereby conferring antimicrobial susceptibility by weakening of the cell envelope barrier in Gram-negative bacteria. A high-throughput whole-cell screening platform was developed to measureEscherichia colienvelope permeability to a β-galactosidase chromogenic substrate. The signal produced by cytoplasmic β-galactosidase-dependent cleavage of the chromogenic substrate was used to determine the degree of envelope permeabilization. The assay was optimized by using known envelope-permeabilizing compounds andE. coligene deletion mutants with impaired envelope integrity. As a proof of concept, a compound library comprising 36 peptides and 45 peptidomimetics was screened, leading to identification of two peptides that substantially increased envelope permeability. Compound 79 reduced significantly (from 8- to 125-fold) the MICs of erythromycin, fusidic acid, novobiocin and rifampin and displayed synergy (fractional inhibitory concentration index, <0.2) with these antibiotics by checkerboard assays in two genetically distinctE. colistrains, including the high-risk multidrug-resistant, CTX-M-15-producing sequence type 131 clone. Notably, in the presence of 0.25 μM of this peptide, both strains were susceptible to rifampin according to the resistance breakpoints (R> 0.5 μg/ml) for Gram-positive bacterial pathogens. The high-throughput screening platform developed in this study can be applied to accelerate the discovery of antimicrobial helper drug candidates and targets that enhance the delivery of existing antibiotics by impairing envelope integrity in Gram-negative bacteria.

Genetic diversity and population structure of Escherichia coli from neighboring small-scale dairy farms

2011 ◽  
Vol 49 (5) ◽  
pp. 693-702 ◽  
Author(s):  
Jesús Andrei Rosales-Castillo ◽  
Ma. Soledad Vázquez-Garcidueñas ◽  
Hugo Álvarez-Hernández ◽  
Omar Chassin-Noria ◽  
Alba Irene Varela-Murillo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genetic Diversity ◽  
Population Structure ◽  
Dairy Farms ◽  
Small Scale ◽  
Small Scale Dairy

Equalizing growth in high-throughput small scale cultivations via precultures operated in fed-batch mode

2009 ◽  
Vol 103 (6) ◽  
pp. 1095-1102 ◽  
Author(s):  
Robert Huber ◽  
Marco Scheidle ◽  
Barbara Dittrich ◽  
Doris Klee ◽  
Jochen Büchs
Keyword(s):  
High Throughput ◽  
Small Scale ◽  
Fed Batch ◽  
Batch Mode
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