Virus Genes

, Volume 30, Issue 1, pp 37–48

Nucleocapsid protein of cell culture-adapted Seoul virus strain 80–39: Analysis of its encoding sequence, expression in yeast and immuno-reactivity

  • Jonas Schmidt
  • Burkhard Jandrig
  • Boris Klempa
  • Kumiko Yoshimatsu
  • Jiro Arikawa
  • Helga Meisel
  • Matthias Niedrig
  • Christian Pitra
  • Detlev H. Krüger
  • Rainer Ulrich
Article

DOI: 10.1007/s11262-004-4580-2

Cite this article as:
Schmidt, J., Jandrig, B., Klempa, B. et al. Virus Genes (2005) 30: 37. doi:10.1007/s11262-004-4580-2

Abstract.

Seoul virus (SEOV) is a hantavirus causing a mild to moderate form of hemorrhagic fever with renal syndrome that is distributed mainly in Asia. The nucleocapsid (N) protein-encoding sequence of SEOV (strain 80-39) was RT-PCR-amplified and cloned into a yeast expression vector containing a galactose-inducible promoter. A survey of the pattern of synonymous codon preferences for a total of 22 N protein-encoding hantavirus genes including 13 of SEOV strains revealed that there is minor variation in codon usage by the same gene in different viral genomes. Introduction of the expression plasmid into yeast Saccharomycescerevisiae resulted in the high-level expression of a hexahistidine-tagged N protein derivative. The nickel-chelation chromatography purified, yeast-expressed SEOV N protein reacted in the immunoblot with a SEOV-specific monoclonal antibody and certain HTNV- and PUUV-cross-reactive monoclonal antibodies. The immunization of a rabbit with the recombinant N protein resulted in the induction of a high-titered antibody response. In ELISA studies, the N protein was able to detect antibodies in sera of experimentally infected laboratory rats and in human anti-hantavirus-positive sera or serum pools of patients from different geographical origin. The yeast-expressed SEOV N protein represents a promising antigen for development of diagnostic tools in serology, sero prevalence studies and vaccine development.

Keywords

codon usageELISAhantavirusmonoclonal antibodiesnucleocapsid proteinSeoul virusyeast expression

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Jonas Schmidt
    • 1
  • Burkhard Jandrig
    • 2
  • Boris Klempa
    • 1
    • 6
  • Kumiko Yoshimatsu
    • 3
  • Jiro Arikawa
    • 3
  • Helga Meisel
    • 1
  • Matthias Niedrig
    • 4
  • Christian Pitra
    • 5
  • Detlev H. Krüger
    • 1
  • Rainer Ulrich
    • 1
    • 7
    • 7
  1. 1.Institute of VirologyCharité Medical SchoolBerlinGermany
  2. 2.Department of Tumor GeneticsMax Delbrueck Center for Molecular MedicineBerlinGermany
  3. 3.Institute for Animal Experimentation, Graduate School of MedicineHokkaido UniversitySapporoJapan
  4. 4.Robert Koch InstituteBerlinGermany
  5. 5.Institute for Zoo and Wildlife ResearchBerlinGermany
  6. 6.Institute of VirologySlovak Academy of SciencesBratislavaSlovakia
  7. 7.Friedrich-Loeffler-InstitutBundesforschungsinstitut für Tiergesundheit, Institut für EpidemiologieWusterhausenGermany