EMD-1905 Experiments and Validation

Residues of the UL25 Protein of Herpes Simplex Virus That Are Required for Its Stable Interaction with Capsids.

Single particle reconstruction
Overview of EMD-1905
Sample name: HSV-1 C-capsid with the UL25 protein labeled with a tandem affinity purification (TAP) tag at amino acid 50.
Organism: Human herpesvirus 1 strain KOS

Map parameters

Minimum density: -7.12
Maximum density: 6.772
Average density: 0.00
Standard deviation: 1.00
Recommended contour level: 0.80 (author)

Sample information

Sample name: HSV-1 C-capsid with the UL25 protein labeled with a tandem affinity purification (TAP) tag at amino acid 50.
Virus: Human herpesvirus 1 strain KOS

Validation information

Experimental information

 

Image processing

Applied symmetry: I
Software: AUTO3DEM
Number of particles: 4865
Reconstruction protocol: Polar Fourier Transform
CTF correction: Phase flip each particle

Imaging

Session
Microscope model: FEI TECNAI F20
Electron source: FIELD EMISSION GUN
Nominal CS (mm): 2.00
Nominal magnification: 50000.0
Defocus max (nm): 3,730.00
Defocus min (nm): 1,250.00
Holder model: GATAN LIQUID NITROGEN
Acquisition
Number of images: 83
Scanner model: NIKON SUPER COOLSCAN 9000
Sampling interval (μm): 6.3499999999999996

Specimen

Preparation
Specimen state: particle
Buffer: 500 mM NaCl, 10 mM Tris, 1 mM EDTA
Vitrification
Apparatus: FEI VITROBOT MARK III
Cryogen: ETHANE
Humidity: 85
Protocol: 6s blot prior to plunging
Details: Vitrification instrument: Vitrobot mark III