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3 ARTICLES PUBLISHED IN JoVE

Biology

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Louise E. Bird¹, Heather Rada¹, Anil Verma¹, Raphael Gasper^2,3, James Birch⁴, Matthew Jennions⁴, Jan Lӧwe³, Isabel Moraes⁴, Raymond J. Owens¹

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.

Biology

Volume Segmentation and Analysis of Biological Materials Using SuRVoS (Super-region Volume Segmentation) Workbench

Michele C. Darrow¹, Imanol Luengo^1,2, Mark Basham¹, Matthew C. Spink¹, Sarah Irvine¹, Andrew P. French², Alun W. Ashton¹, Elizabeth M.H. Duke¹

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.

Biochemistry

Analysis of SEC-SAXS data via EFA deconvolution and Scatter

Mark D Tully^*1, Nicolas Tarbouriech², Robert P Rambo³, 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.