High yields of stable and highly pure nucleocapsid proteins of different hantaviruses can be generated in the yeast

High yields of stable and highly pure nucleocapsid proteins of different hantaviruses can be generated in the yeast
5 August 2004
Received 16 October 2003; Revised 14 April 2004; accepted 30 April 2004. Available online 19 June 2004.
Aura Razanskiene, , a, Jonas Schmidtb, Astrid Geldmacherb, 1, Andreas Ritzib, Matthias Niedrigc, ?ke Lundkvistd, e, Detlev H. Kr?gerb, Helga Meiselb, Kestutis Sasnauskasa and Rainer Ulrichb, 2, 3
Journal of Biotechnology
Volume 111, Issue 3 , 5 August 2004, Pages 319-333
ScienceDirect
a Institute of Biotechnology, V. Graiciuno 8, LT-2028, Vilnius, Lithuania
b Institute of Virology, Charit? Medical School, D-10098, Berlin, Germany
c Robert Koch Institute, Nordufer 20, D-13353 Berlin, Germany
d Microbiology and Tumor Biology Center, Karolinska Institutet, S-171 77, Stockholm, Sweden
e Swedish Institute for Infectious Disease Control, S-171 82, Stockholm, Sweden
Abstract
Recently, the high-level expression of authentic and hexahistidine (His)-tagged Puumala (strain Vranica/H?lln?s) hantavirus nucleocapsid protein derivatives in the yeast Saccharomyces cerevisiae has been reported [Dargeviciute et al., Vaccine, 20 (2002) 3523?3531]. Here we describe the expression of His-tagged nucleocapsid proteins of other Puumala virus strains (Sotkamo, Kazan) as well as Dobrava (strains Slovenia and Slovakia) and Hantaan (strain Fojnica) hantaviruses using the same system. All nucleocapsid proteins were expressed in the yeast S. cerevisiae at high levels. The nucleocapsid proteins can be easily purified by nickel chelate chromatography; the yield for all nucleocapsid proteins ranged from 0.5 to 1.5 mg per g wet weight of yeast cells. In general, long-term storage of all nucleocapsid proteins without degradation can be obtained by storage in PBS at -20 ?C or lyophilization. The nucleocapsid protein of Puumala virus (strain Vranica/H?lln?s) was demonstrated to contain only traces of less than 10 pg nucleic acid contamination per 100 g of protein. The yeast-expressed nucleocapsid proteins of Hantaan, Puumala and Dobrava viruses described here represent useful tools for serological hantavirus diagnostics and for vaccine development.
Author Keywords: Hantavirus; Recombinant nucleocapsid protein; Yeast; Antigenicity; Immunogenicity
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