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Expression and X-Ray Structural Determination of the Nucleoprotein of Lassa Fever Virus

Part of the Methods in Molecular Biology book series (MIMB,volume 1604)

Abstract

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.

Key words

  • Lassa virus
  • Nucleoprotein
  • X-ray structure
  • Large-scale expression
  • Arenavirus

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Acknowledgments

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.

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Correspondence to Changjiang Dong or Hinh Ly .

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Qi, X., Wang, W., Dong, H., Liang, Y., Dong, C., Ly, H. (2018). Expression and X-Ray Structural Determination of the Nucleoprotein of Lassa Fever Virus. In: Salvato, M. (eds) Hemorrhagic Fever Viruses. Methods in Molecular Biology, vol 1604. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6981-4_12

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  • DOI: https://doi.org/10.1007/978-1-4939-6981-4_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6980-7

  • Online ISBN: 978-1-4939-6981-4

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