University of Wroclaw

4 ARTICLES PUBLISHED IN JoVE

Biology

Live-Cell Imaging of the Life Cycle of Bacterial Predator Bdellovibrio bacteriovorus using Time-Lapse Fluorescence Microscopy

Łukasz Makowski¹, Damian Trojanowski¹, Jolanta Zakrzewska-Czerwińska¹

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.

Biology

Demonstration of Heterologous Complexes formed by Golgi-Resident Type III Membrane Proteins using Split Luciferase Complementation Assay

Wojciech Wiertelak¹, Mariusz Olczak¹, Dorota Maszczak-Seneczko¹

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.

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 Blaszczak³, Nicolas Bertaux⁴, Marcin Łukaszewicz², Didier Marguet¹, Tomasz Trombik²

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.

Biochemistry

Preparation of Site-Specific Cytotoxic Protein Conjugates via Maleimide-thiol Chemistry and Sortase A-Mediated Ligation

Mateusz Adam Krzyscik^1,2, Aleksandra Sokolowska-Wedzina¹, Karolina Jendryczko¹, Marta Pozniak¹, Daria Nawrocka¹, Natalia Porebska¹, Malgorzata Zakrzewska¹, Jacek Otlewski¹, Anna Szlachcic¹, Lukasz Opalinski¹

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.