Accedi

Bioeconomy Science Center (BioSC)

3 ARTICLES PUBLISHED IN JoVE

Bioengineering

Microfluidic Picoliter Bioreactor for Microbial Single-cell Analysis: Fabrication, System Setup, and Operation

Alexander Gruenberger¹, Christopher Probst¹, Antonia Heyer¹, Wolfgang Wiechert¹, Julia Frunzke¹, Dietrich Kohlheyer¹

¹Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Juelich GmbH

In this protocol the fabrication, setup and basic operation of a microfluidic picoliter bioreactor (PLBR) for single-cell analysis of prokaryotic microorganisms is introduced. Industrially relevant microorganisms were analyzed as proof of principle allowing insights into growth rate, morphology, and phenotypic heterogeneity over certain time periods, hardly possible with conventional methods.

Genetics

Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions

Kristin Bösch^*1,2, Lamprinos Frantzeskakis^*1, Miroslav Vraneš³, Jörg Kämper³, Kerstin Schipper^1,2, Vera Göhre^1,2,4

¹Institute for Microbiology, Heinrich-Heine University Düsseldorf, ²Bioeconomy Science Center (BioSC), ³Department of Genetics, Institute of Applied Biosciences, Karlsruhe Institute of Technology, ⁴Cluster of Excellence in Plant Sciences (CEPLAS), Heinrich-Heine University Düsseldorf

We describe a robust gene replacement strategy to genetically manipulate the smut fungus Ustilago maydis. This protocol explains how to generate deletion mutants to investigate infection phenotypes. It can be extended to modify genes in any desired way, e.g., by adding a sequence encoding a fluorescent protein tag.

Bioengineering

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology

Johannes Hemmerich^*1,2, Lars Freier^*1,2, Wolfgang Wiechert^1,2,3, Eric von Lieres^1,2, Marco Oldiges^1,2,4

¹IBG-1: Biotechnology, Forschungszentrum Jülich, ²Research Center Jülich, Bioeconomy Science Center (BioSC), ³Computational Systems Biotechnology (AVT.CSB), RWTH Aachen University, ⁴Institute for Biotechnology, RWTH Aachen University

This manuscript describes a generic approach for tailor-made design of microbial cultivation media. This is enabled by an iterative workflow combining Kriging-based experimental design and microbioreactor technology for sufficient cultivation throughput, which is supported by lab robotics to increase reliability and speed in liquid handling media preparation.