Author: Wilton T. Snead; Wade F. Zeno; Grace Kago; Ryan W. Perkins; J Blair Richter; Chi Zhao; Eileen M. Lafer; Jeanne C. Stachowiak
Title: BAR scaffolds drive membrane fission by crowding disordered domains Document date: 2018_3_4
ID: drqseaaa_45
Snippet: The pGex6P bacterial expression vector containing full-length human amphiphysin (residues 2-695) was generously provided by the Baumgart Lab (University of Pennsylvania). The N-BAR domain of human amphiphysin (residues 2-242) was cloned into the pGex4T2 bacterial expression vector using BamHI and EcoRI restriction sites. The C-terminal domain of human amphiphysin lacking the SH3 domain (Amph CTD DSH3, residues 240-622) with N-terminal GST and 6hi.....
Document: The pGex6P bacterial expression vector containing full-length human amphiphysin (residues 2-695) was generously provided by the Baumgart Lab (University of Pennsylvania). The N-BAR domain of human amphiphysin (residues 2-242) was cloned into the pGex4T2 bacterial expression vector using BamHI and EcoRI restriction sites. The C-terminal domain of human amphiphysin lacking the SH3 domain (Amph CTD DSH3, residues 240-622) with N-terminal GST and 6his tags was cloned using a previously-generated plasmid template, GST-6his-AP180 CTD in pGex4T2 . AP180 CTD was excised from the template using SalI and XhoI restriction sites and the Amph CTD ∆SH3 insert was ligated in using the same SalI and XhoI sites. The N-BAR domain of human amphiphysin fused to the C-terminal domain of rat epsin1 (N-BAR-epsin CTD, residues 144-575 of rat epsin1) was cloned by first ligating in the N-BAR All rights reserved. No reuse allowed without permission.
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