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Virus Genes

, Volume 34, Issue 2, pp 199–204 | Cite as

Expression of porcine circovirus 2 ORF2 gene requires codon optimized E. coli cells

  • Maria Trundova
  • Vladimir Celer
Original Article

Abstract

Expression and purification of whole and nuclear localization signal (NLS) deleted ORF2 capsid protein of porcine circovirus 2 (PCV2) is demonstrated in the present study. Gene coding for both protein forms were cloned into pDest17 vector and expressed in BL21 (DE3)AI cells and in BL21-CodonPlus (DE3)-RIPL E. coli cells. The later cells were used to overcome difficulties with the heterologous expression of viral proteins in prokaryotic systems. Whole 30 kDa recombinant ORF2 protein was successfully expressed in BL21-CodonPlus (DE3)-RIPL cells only, 3 mg of pure protein was consistently obtained per liter of bacterial culture. NLS deleted ORF2 protein was expressed in both cell types. Resulting proteins reacted with PCV2 positive swine serum in immunofluorescent test and immunoblot.

Keywords

PMWS PCV2 Protein expression Nuclear localization signal 

Notes

Acknowledgment

This work was supported by the Grant Agency of the Czech Republic, Grant No. 524/06/1640.

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Copyright information

© Springer Science+Business Media, LLC 2006

Authors and Affiliations

  1. 1.Institute of Microbiology and Immunology, Faculty of Veterinary MedicineVeterinary and Pharmaceutical University BrnoBrnoCzech Republic

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