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From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins
 

From Constructs to Crystals – Towards Structure Determination of β-barrel Outer Membrane Proteins

Article DOI: 10.3791/53245-v • 09:55 min • July 4th, 2016
July 4th, 2016 •
1, 2, 2,3, 2,3, 2
1Department of Biological Sciences, Markey Center for Structural Biology, Purdue University, 2National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health, 3National Institute of General Medical Sciences (NIGMS), National Institutes of Health

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β-barrel outer membrane proteins (OMPs) serve many functions within the outer membranes of Gram-negative bacteria, mitochondria, and chloroplasts. Here, we hope to alleviate a known bottleneck in structural studies by presenting protocols for the production of β-barrel OMPs in sufficient quantities for structure determination by X-ray crystallography or NMR spectroscopy.

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Constructs Crystals Structure Determination Beta-barrel Outer Membrane Proteins Milligram Quantities Crystallization Defraction Membrane Protein Folding Transport Cell Membranes Detergents Purification Expression Vector Codon Optimized Target Outer Membrane Protein OMP Gene In Vivo Expression Bacteria BL21(DE3) Chemically Competent Cells Incubation Heat Pulse Water Bath SOC Media
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