Hpylori_dFliM

Description

Technical info

Tilt Series Date: 2019-11-24

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: 120.0 eV/Ų, defocus: -8.0 μm, magnification: 19500x.

Microscope: Caltech Titan Krios

Acquisition Software: Serial EM

Upload Method: pipeline

Processing Software Used: None

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 FliM 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_dfliM_058_af.mrc, Tilt Series (Pixel Size 0.449 nm), 1.9 GB