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3 ARTICLES PUBLISHED IN JoVE

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Biology

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
Louise E. Bird 1, Heather Rada 1, Anil Verma 1, Raphael Gasper 2,3, James Birch 4, Matthew Jennions 4, Jan Lӧwe 3, Isabel Moraes 4, Raymond J. Owens 1
1Oxford Protein Production Facility, Research Complex at Harwell, 2Protein Crystallography Group, Ruhr University, 3MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, 4Membrane Protein Laboratory, Diamond Light Source

A streamlined approach to screening for the expression of recombinant membrane proteins in Escherichia coli based on fusion to green fluorescent protein is presented.

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Biology

Volume Segmentation and Analysis of Biological Materials Using SuRVoS (Super-region Volume Segmentation) Workbench
Michele C. Darrow 1, Imanol Luengo 1,2, Mark Basham 1, Matthew C. Spink 1, Sarah Irvine 1, Andrew P. French 2, Alun W. Ashton 1, Elizabeth M.H. Duke 1
1Science Division, Harwell Science and Innovation Campus, Diamond Light Source, 2School of Computer Science, University of Nottingham

Segmentation of three-dimensional data from many imaging techniques is a major bottleneck in analysis of complex biological systems. Here, we describe the use of SuRVoS Workbench to semi-automatically segment volumetric data at various length-scales using example datasets from cryo-electron tomography, cryo soft X-ray tomography, and phase contrast X-ray tomography techniques.

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Biochemistry

Analysis of SEC-SAXS data via EFA deconvolution and Scatter
Mark D Tully *1, Nicolas Tarbouriech 2, Robert P Rambo 3, Stephanie Hutin *4
1European Synchrotron Radiation Facility Structural Biology Group, Structural Biology Group, 2Institut de Biologie Structurale, University Grenoble Alpes, CEA, CNRS, 3Diamond Light Source, 4Laboratoire de Physiologie Cellulaire and Végétale, Université Grenoble Alpes/CNRS/CEA/INRA/BIG

SEC-BioSAXS measurements of biological macromolecules are a standard approach for determining solution structure of macromolecules and their complexes. Here, we analyze SEC-BioSAXS data from two types of commonly encountered SEC traces—chromatograms with fully resolved and partially resolved peaks. We demonstrate the analysis and deconvolution using scatter and BioXTAS RAW.

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