lip10-2 (+ megacomplex)

Description

Technical info

Tilt Series Date: 2020-09-29

Data Taken By: William Nicolas

Species / Specimen: Escherichia coli

Strain: PK2, BL21 dHisdHA RQABpCDF+EFGpRSF

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 2°, constant angular increment, dosage: 56.0 eV/Ų, defocus: -8.0 μm, magnification: 11000x.

Microscope: IECB Talos Artica

Acquisition Software: Serial EM

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: Petya Krasteva (head of the SBB lab at the IECB, Bordeaux° and Wiem . Abidi (graduate student) did the preps and helped with acquisition

Purification / Growth Conditions / Treatment: BL21 cells were transformed with QAB - pCDF plasmid and EFG - pRSF . plasmid to express the inner-membrane macro-complex.. . Selection done with Kanamycin and Streptomycin.. . Induction of expression ON done with IPTG. . Spheroplasts are generated from the BL21 non transformed cells cells.. Spheroplasts are processed through a high-pressure homogenizer and . further purification by ultracentrifugation of the membrane . fractions. . Liposomes are then generated from the membrane fractions by . processing through Potter and sonicator..

Sample Preparation: Fiducials were prepared my method (see lab archive). . Grids: Quantifoil Cu NH2 R2/2. . Glowdischarged 3mA - 2min. BT3s - BF0. 4uL pipetted on the grids

Attached Files

lip10-2-sq1-tg8.mrc, Tilt Series (Pixel Size 0.39 nm), 1.7 GB

lip10-2-sq1-tg8_full.rec, Reconstruction (Pixel Size 0.78 nm), 1.8 GB