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University of Wroclaw

4 ARTICLES PUBLISHED IN JoVE

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Biology

Live-Cell Imaging of the Life Cycle of Bacterial Predator Bdellovibrio bacteriovorus using Time-Lapse Fluorescence Microscopy
Łukasz Makowski 1, Damian Trojanowski 1, Jolanta Zakrzewska-Czerwińska 1
1Department of Molecular Microbiology, Faculty of Biotechnology, University of Wroclaw

Presented here is a protocol that describes monitoring of the complete life cycle of predatory bacterium Bdellovibrio bacteriovorus using time-lapse fluorescence microscopy in combination with an agarose pad and cell-imaging dishes.

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Biology

Demonstration of Heterologous Complexes formed by Golgi-Resident Type III Membrane Proteins using Split Luciferase Complementation Assay
Wojciech Wiertelak 1, Mariusz Olczak 1, Dorota Maszczak-Seneczko 1
1Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw

The protocol presented here is intended to demonstrate the occurrence of heterologous interactions between Golgi-resident type III membrane proteins with cytoplasmically exposed N- and/or C-termini in live mammalian cells using the most recent variant of the split luciferase complementation assay.

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Biochemistry

Spot Variation Fluorescence Correlation Spectroscopy for Analysis of Molecular Diffusion at the Plasma Membrane of Living Cells
Sebastien Mailfert *1, Karolina Wojtowicz *2, Sophie Brustlein *1, Ewa Blaszczak 3, Nicolas Bertaux 4, Marcin Łukaszewicz 2, Didier Marguet 1, Tomasz Trombik 2
1CNRS, Inserm, Centre d'Immunologie de Marseille-Luminy, Aix Marseille Univ, 2Faculty of Biotechnology, University of Wroclaw, 3Faculty of Biological Sciences, University of Wroclaw, 4CNRS, Centrale Marseille, Institut Fresnel, Aix Marseille Univ

This article aims to present a protocol on how to build a spot variation Fluorescence Correlation Spectroscopy (svFCS) microscope to measure molecular diffusion at the plasma membrane of living cells.

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Biochemistry

Preparation of Site-Specific Cytotoxic Protein Conjugates via Maleimide-thiol Chemistry and Sortase A-Mediated Ligation
Mateusz Adam Krzyscik 1,2, Aleksandra Sokolowska-Wedzina 1, Karolina Jendryczko 1, Marta Pozniak 1, Daria Nawrocka 1, Natalia Porebska 1, Malgorzata Zakrzewska 1, Jacek Otlewski 1, Anna Szlachcic 1, Lukasz Opalinski 1
1Department of Protein Engineering, Faculty of Biotechnology, University of Wroclaw, 2Department of Protein Biotechnology, Faculty of Biotechnology, University of Wroclaw

Here we provide detailed protocols for a site-specific labeling of proteins with cytotoxic drugs using maleimide-thiol reaction and sortase A-mediated ligation.

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