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Extracellular vesicles as a platform to study cell-surface membrane proteins

Methods. 2020; 
Delauzun V, Amigues B, Gaubert A, Leone P, Grange M, Gauthier L, Roussel A
Products/Services Used Details Operation
Gene Synthesis … Zaire Ebola virus glycoprotein Δ934-1389 cDNA (Zebov GP) was ordered from Genscript company
as a synthetic gene cloned into the pcDNA31 vector (Thermo Fisher #V79020) This cDNA codes
for the envelope's glycoprotein without its mucin domain [24] …
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摘要

Producing intact recombinant membrane proteins for structural studies is an inherently challenging task due to their requirement for a cell-lipid environment. Most of the procedures developed involve isolating the protein by solubilization with detergent and further reconstitutions into artificial membranes. These procedures are highly time consuming and suffer from further drawbacks, including low yields and high cost. We describe here an alternative method for rapidly obtaining recombinant cell-surface membrane proteins displayed on extracellular vesicles (EVs) derived from cells in culture. Interaction between these membrane proteins and ligands can be analyzed directly on EVs. Moreover, EVs can also be used... More

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