H_pylori_dFliO_GC

Description

Technical info

Tilt Series Date: 2019-10-25

Data Taken By: Mohammed Kaplan

Species / Specimen: Helicobacter pylori

Strain: 26695

Tilt Series Settings: Single Axis, tilt range: (-60.0°, 60.0°), step: 3°, constant angular increment, dosage: 130.0 eV/Ų, defocus: -6.0 μm, magnification: 19500x.

Microscope: Caltech Titan Krios

Acquisition Software: Serial EM

Upload Method: pipeline

Processing Software Used: Raptor

Collaborators and Roles: I received these cells from Carrie Schaffer from the university of Kentucky. These are wild . . type cells but they don't express complete FliP protein. Therefore, they don't assemble full . . flagella. We knocked out FliO protein of the flagellar motor to study the early assembly states . of flagellar motor.

Purification / Growth Conditions / Treatment: I grew the cells on blood agar plates from -80 stock at 37C with 5% CO2. the cells were grown . . for 48 hours and then spread on a new plate with a cotton swab and left to grow for 24 hours. . . The grids were prepared from both batches (48 hours, and 24 hours, separately).

Sample Preparation: The samples were prepared with Vitrobot IV. Blotting conditions were either: blot force 6, blot . . time 3, drain time 1 OR blot force -3, blot time 4, drain time 1, 100% humidity. Cu grids. 10 nm . . gold.

Attached Files

Hpylori_FliO_049_af.mrc, Tilt Series (Pixel Size 0.449 nm), 3.9 GB

Hpylori_FliO_049_af_full.rec, Reconstruction (Pixel Size 1.796 nm), 737.3 MB