Back to Search
Start Over
Transient expression of the influenza A virus PB1-F2 protein using a plum pox virus-based vector in Nicotiana benthamiana
- Source :
- Acta virologica. 58(3)
- Publication Year :
- 2014
-
Abstract
- PB1-F2 protein of influenza A virus (IAV) was cloned in a plum pox virus (PPV) genome-based vector and attempts to express it in biolistically transfected Nicotiana benthamiana plants were performed. The vector-insert construct replicated in infected plants properly and was stable during repeated passage by mechanical inoculation, as demonstrated by disease symptoms and immunoblot detection of PPV capsid protein, while PB1-F2-specific band was more faint. We showed that it was due its low solubility. Modification of sample preparation (denaturation/solubilization preceding the centrifugation of cell debris) led to substantial signal enhancement. Maximal level of PB1-F2 expression in plants was observed 12 days post inoculation (dpi). Only 1% SDS properly solubilized the protein, other detergents were much less efficient. Solubilization with 8M urea released approximately 50% of PB1-F2 from the plant tissues, thus the treatment with this removable chaotropic agent may be a good starting point for the purification of the protein for eventual functional studies in the future.
- Subjects :
- viruses
Genetic Vectors
Nicotiana benthamiana
Gene Expression
medicine.disease_cause
Protein Engineering
Viral Proteins
Virology
Tobacco
Influenza A virus
medicine
Centrifugation
Denaturation (biochemistry)
biology
Chemistry
Inoculation
fungi
virus diseases
General Medicine
Transfection
biology.organism_classification
Chaotropic agent
Infectious Diseases
Capsid
Plum Pox Virus
Subjects
Details
- ISSN :
- 0001723X
- Volume :
- 58
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Acta virologica
- Accession number :
- edsair.doi.dedup.....38e4da0f364e79a2d3ed7c28e83e4f50