We describe methods to express the nucleoprotein (NP) of Lassa fever virus (LASV) in E. coli, to purify and crystallize it using the sitting-drop vapor diffusion method. The crystals were screened using Rigaku micro-007 X-ray generator and a dataset was collected at a resolution of 2.36 Å. The crystals belong to space group P3, with the unit cell parameters a = b = 176.35 Å, c = 56.40 Å, α = β = 90°, and γ = 120°. Using the X-ray diffraction method, we constructed a three-dimensional structure of the LASV NP that should aid in the development of novel therapeutic strategies against this virus, for which vaccine and effective treatment modalities are currently unavailable.
|Original language||English (US)|
|Title of host publication||Methods in Molecular Biology|
|Publisher||Humana Press Inc.|
|Number of pages||10|
|State||Published - 2018|
|Name||Methods in Molecular Biology|
Bibliographical noteFunding Information:
We thank Dr. Louise Major for providing the pLou3 plasmid, Dr. Catherine Botting for assistance with mass spectroscopy, Dr. Zaitsev Slava for helping with data collection, and Dr. Bjoern Meyer for prompt editorial assistance. This work was supported in part by the NIAID/NIH through the new-direction awards mechanism of the SERCEB grant (U54-AI057157) to YL and HL, by the NIAID/NIH R01 AI083409 to YL, R01 AI093580 and R56 AI091805 to HL, and the Medical Research Council grant (G1100110/1) to CJD.
© Springer Science+Business Media LLC 2018.
- Large-scale expression
- Lassa virus
- X-ray structure