Expression and Ni-NTA-Agarose Purification of Recombinant Hepatitis C Virus E2Ectodomain Produced in a Baculovirus Expression System
| Autor: | Francisco Gavilanes, Julián Gómez-Gutiérrez, Belén Yélamos, Mar Rodríguez-Rodríguez |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Strategy and Management Hepatitis C virus medicine.disease_cause Industrial and Manufacturing Engineering Virus law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Affinity chromatography law medicine Extracellular Methods Article chemistry.chemical_classification Mechanical Engineering Metals and Alloys Amino acid 030104 developmental biology Ectodomain chemistry Biochemistry Recombinant DNA Agarose 030217 neurology & neurosurgery |
| Zdroj: | Bio Protoc |
| ISSN: | 2331-8325 |
| Popis: | In this protocol, we describe the production and purification of the ectodomain of the E2(661) envelope protein (amino acids 384-661) of the Hepatitis C virus, which plays a fundamental role in the entry of the virus into the host cell. This protein has been expressed in both prokaryotic and eukaryotic systems but in small quantities or without native protein characteristics. In our case, we use the Baculovirus expression system in insect cells. E2(661) is secreted into the extracellular medium and purified by means of affinity chromatography a Ni-NTA-column because the protein has a tag of six histidines at its amino terminal end. The purified protein possesses a native-like conformation and it is produced in large quantities, around 5-6 mg per liter. |
| Databáze: | OpenAIRE |
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