JoVE Journal

Chemistry

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Nanyang Technological University

מתוך בונה קריסטלים - לקראת מבנה קביעת β-חבית החיצון ממברנה חלבונים

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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Chapters in this video

0:05

Title

0:46

In Vivo Membrane-targeted Expression

3:09

Isolation from Membrane Fractions

4:41

Purification Using Immobilized Metal-affinity Chromatography (IMAC), Ion-exchange, and Gel Filtration

7:09

Heat Modifiability Assay

8:35

Results: Crystals to Structure Determination for the Putative Vaccine Target YiuR from Yersinia pestis

9:29

Conclusion

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