Heterologous expression and isolation of influenza A virus nuclear export protein NEP
- Авторы: Golovko A.1, Koroleva O.1, Drutsa V.2
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Учреждения:
- Lomonosov Moscow State University
- Belozersky Institute of Physico-Chemical Biology
- Выпуск: Том 82, № 12 (2017)
- Страницы: 1529-1537
- Раздел: Regular Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151542
- DOI: https://doi.org/10.1134/S0006297917120124
- ID: 151542
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Аннотация
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.
Об авторах
A. Golovko
Lomonosov Moscow State University
Email: koroleva@genebee.msu.ru
Россия, Moscow, 119991
O. Koroleva
Lomonosov Moscow State University
Автор, ответственный за переписку.
Email: koroleva@genebee.msu.ru
Россия, Moscow, 119991
V. Drutsa
Belozersky Institute of Physico-Chemical Biology
Email: koroleva@genebee.msu.ru
Россия, Moscow, 11991