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Published on: October 9, 2011
Expression of the gp150 maedi visna virus envelope precursor protein by mammalian expression vectors
Christophe Fraisier1, Hallgrimur Arnarson, Cyril Barbezange
1Unité des Rickettsies, CNRS UMR 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 27 Boulevard Jean Moulin, 13385 Marseille Cedex 05, France.
Abstract:
There are very few previous reports of expression of native full-length maedi visna virus (MVV) Env gp150 protein in the literature. Therefore the use of different plasmid and viral expression vectors to obtain full-length gp150 was investigated. A mammalian expression plasmid, pN3-Env, was constructed containing the MVV env gene encoding the precursor protein gp150 Env. The functionality of the recombinant plasmid was tested for expression in HEK293 cells. A recombinant modified vaccinia Ankara virus, MVA-Env, with expression detected in avian cells was also made. The expression of the MVV gp150 Env precursor protein was shown for the first time upon transfection of the eukaryotic HEK293 cells by the pN3-Env plasmid DNA as demonstrated by Western blot analysis. These plasmid or viral expression vectors are of potential use in MVV vaccines.
Insights
Researchers explored novel methods for expressing the full-length Maedi Visna Virus (MVV) Env gp150 protein. They successfully demonstrated its expression using plasmid and viral vectors, paving the way for potential MVV vaccines.
Area of Science:
- Virology
- Molecular Biology
- Vaccine Development
Background:
- Limited reports exist on expressing the native full-length Maedi Visna Virus (MVV) Env gp150 protein.
- Developing effective expression systems is crucial for advancing MVV research and vaccine strategies.
Purpose of the Study:
- To investigate the efficacy of plasmid and viral expression vectors for producing full-length MVV Env gp150 protein.
- To establish a reliable method for expressing the MVV gp150 Env precursor protein.
Main Methods:
- Construction of a mammalian expression plasmid (pN3-Env) containing the MVV env gene.
- Testing expression in HEK293 eukaryotic cells via plasmid transfection.
- Development of a recombinant modified vaccinia Ankara virus (MVA-Env) for expression in avian cells.
- Western blot analysis to confirm protein expression.
Main Results:
- Successfully demonstrated the expression of the MVV gp150 Env precursor protein in HEK293 cells using the pN3-Env plasmid.
- Confirmed expression of the MVV Env protein in avian cells using the MVA-Env viral vector.
- This marks the first reported instance of expressing the native full-length MVV gp150 Env precursor protein via plasmid transfection.
Conclusions:
- The developed plasmid and viral expression vectors are effective for producing the MVV gp150 Env precursor protein.
- These expression systems hold significant potential for the development of Maedi Visna Virus vaccines.
