Q73140094 Dotaz Zobrazit nápovědu
Transient expression of foreign genes based on plant viral vectors is a suitable system for the production of relevant immunogens that can be used for the development of a new generation of vaccines against a variety of infectious diseases. In the present study the epitope derived from HPV-16 L2 minor capsid protein (amino acids 108-120) was expressed from Potato virus X (PVX)-based vector pGR106 as N- or C-terminal fusion with the PVX coat protein (PVX CP) in transgenic Nicotiana benthamiana plants. The fusion protein L2 108-120-PVX CP was successfully expressed in plants at a level of 170 mg/kg of fresh leaf tissue. The C-terminal fusion protein PVX CP- L2 108-120 was expressed using mutated vector sequence to avoid homologous recombination at a level of 8 mg/kg of fresh leaf tissue. Immunogenicity of L2 108-120-PVX CP virus-like particles was tested after immunization of mice by subcutaneous injection or tattoo administration. In animal sera the antibodies against the PVX CP and the L2 108-120 epitope were found after both methods of vaccine delivery.
- MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- ELISA MeSH
- epitopy genetika metabolismus MeSH
- genetické vektory genetika MeSH
- geneticky modifikované rostliny MeSH
- imunizace MeSH
- klonování DNA MeSH
- lidé MeSH
- listy rostlin metabolismus MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- oligonukleotidy genetika MeSH
- onkogenní proteiny virové metabolismus MeSH
- protilátky virové krev MeSH
- rekombinantní fúzní proteiny imunologie metabolismus MeSH
- tabák metabolismus MeSH
- transmisní elektronová mikroskopie MeSH
- virion imunologie MeSH
- virové plášťové proteiny metabolismus MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The gene encoding the coat protein (CP) of a potato virus Y (PVY) was cloned into expression vector pMPM-A4Omega. PVY CP was expressed in Escherichia coli and the purified recombinant protein was used for raising rabbit polyclonal antibodies. The sera and antibodies were tested for the detection of PVY in the laboratory host Nicotiana tabacum cv. Petit Havana SR1 and in various cultivars of the natural host Solanum tuberosum by ELISA as well as by Western blots. The antibodies can be used for the detection of the whole strain spectrum of PVY by indirect plate trapped antigen ELISA and Western blot, but not by double antigen sandwich ELISA.
- MeSH
- ELISA MeSH
- králíci MeSH
- monoklonální protilátky biosyntéza MeSH
- nemoci rostlin virologie MeSH
- Potyvirus imunologie MeSH
- protilátky virové biosyntéza MeSH
- rekombinantní proteiny imunologie MeSH
- Solanum tuberosum virologie MeSH
- tabák virologie MeSH
- virové plášťové proteiny imunologie MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH