BMB Reports
- 제33권4호
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- Pages.294-299
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- 2000
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- 1976-670X(eISSN)
The Two-Component Protease NS2B-NS3 of Dengue Virus Type 2: Cloning, Expression in Escherichia coli and Purification of the NS2B, NS3(pro) and NS2B-NS3 Proteins
- Champreda, Veerawat (Institute of Molecular Biology and Genetics, Mahidol University) ;
- Khumthong, Rabuesak (Institute of Molecular Biology and Genetics, Mahidol University) ;
- Subsin, Benchamas (Institute of Molecular Biology and Genetics, Mahidol University) ;
- Angsuthanasombat, Chanan (Institute of Molecular Biology and Genetics, Mahidol University) ;
- Panyim, Sakol (Institute of Molecular Biology and Genetics, Mahidol University) ;
- Katzenmeier, Gerd (Institute of Molecular Biology and Genetics, Mahidol University)
- 투고 : 2000.03.24
- 심사 : 2000.05.08
- 발행 : 2000.07.31
초록
Proteolytic processing of the dengue virus serotype 2 polyprotein precursor is catalyzed by a host signal peptidase and a virus encoded two-component protease consisting of the nonstructural proteins, NS2B and NS3. We expressed in Escherichia coli the NS2B, NS3(pro) and NS2B-NS3 proteins from the dengue virus type 2 strain 16681 as N-terminal fusions with a hexahistidine affinity tag under the control of the inducible trc promoter. All fusion proteins were purified to >90% purity by detergent extraction of inclusion bodies and a single step metal chelate chromatography. Proteins were refolded on-column and recovered with yields of 0.5, 6.0 and 1.0 mg/l of E. coli culture that was grown to
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