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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Papaya Ringspot Virus Coat Protein Gene for Antigen Presentation in Escherichia coli

  • Chatchen, Supawat (Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus) ;
  • Juricek, Mila (Institute of Experimental Botany, Czech Academy of Sciences) ;
  • Rueda, Paloma (Ingenasa, Hermanos Garcia Noblejas, Inmunologia y Genetica Apl. S.A. (INGENASA)) ;
  • Kertbundit, Sunee (Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus)
  • Received : 2005.05.18
  • Accepted : 2005.08.19
  • Published : 2006.01.31
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Abstract

The coat protein (CP) of Papaya ringspot virus (PRSV) was analyzed for presentation of the antigenic peptide of animal virus, Canine parvovirus (CPV), in Escherichia coli (E. coli). The 45 nucleotides fragment coding for the 15-aa peptide epitope of the CPV-VP2 protein was either inserted into the PRSV-cp gene at the 5', 3' ends, both 5' and 3' ends or substituted into the 3' end of the PRSV cp gene. Each of the chimeric PRSV cp genes was cloned into the pRSET B vector under the control of the T7 promoter and transformed into E. coli. The recombinant coat proteins expressed from different chimeric PRSV-cp genes were purified and intraperitoneally injected into mice. All of the recombinant coat proteins showed strong immunogenicity and stimulate mice immune response. The recombinant coat proteins containing the CPV epitope insertion at the C terminus and at both N and C termini elicited ten times higher specific antisera in immunized mice compared with the other two recombinant coat proteins which contain the CPV epitope insertion at the N terminus and substitution at the C terminus.

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References

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