Biochemistry (Moscow)

, Volume 82, Issue 12, pp 1529–1537 | Cite as

Heterologous expression and isolation of influenza A virus nuclear export protein NEP

  • A. O. Golovko
  • O. N. KorolevaEmail author
  • V. L. Drutsa
Regular Article


Influenza A virus nuclear export protein NEP (NS2, 14.4 kDa) plays a key role in various steps of the virus life cycle. Highly purified protein preparations are required for structural and functional studies. In this study, we designed a series of Escherichia coli plasmid constructs for highly efficient expression of the NEP gene under control of the constitutive trp promoter. An efficient method for extraction of NEP from inclusion bodies based on dodecyl sulfate treatment was developed. Preparations of purified NEP with either N-or C-terminal (His)6-tag were obtained using Ni-NTA agarose affinity chromatography with yield of more than 20 mg per liter of culture. According to CD data, the secondary structure of the proteins matched that of natural NEP. A high propensity of NEP to aggregate over a wide range of conditions was observed.


influenza A virus nuclear export protein (NEP) affinity chromatography protein aggregation 



amino acid residue


base pairs




nuclear export protein


NEP with C-terminal (His)6-tag


NEP with N-terminal (His)6-tag


polymerase chain reaction


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Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • A. O. Golovko
    • 1
  • O. N. Koroleva
    • 2
    Email author
  • V. L. Drutsa
    • 3
  1. 1.Lomonosov Moscow State UniversityFaculty of Bioengineering and BioinformaticsMoscowRussia
  2. 2.Lomonosov Moscow State UniversityFaculty of ChemistryMoscowRussia
  3. 3.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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