Abstract
Orthopoxviruses (OPVs) have recently received increasing attention because of their potential use in bioterrorism and the occurrence of zoonotic OPV outbreaks, highlighting the need for the development of safe and cost-effective vaccines against smallpox and related viruses. In this respect, the production of subunit protein-based vaccines in transgenic plants is an attractive approach. For this purpose, the A27L immunogenic protein of vaccinia virus was expressed in tobacco using stable transformation of the nuclear or plastid genome. The vaccinia virus protein was expressed in the stroma of transplastomic plants in soluble form and accumulated to about 18% of total soluble protein (equivalent to approximately 1.7 mg/g fresh weight). This level of A27L accumulation was 500-fold higher than that in nuclear transformed plants, and did not decline during leaf development. Transplastomic plants showed a partial reduction in growth and were chlorotic, but reached maturity and set fertile seeds. Analysis by immunofluorescence microscopy indicated altered chlorophyll distribution. Chloroplast-synthesized A27L formed oligomers, suggesting correct folding and quaternary structure, and was recognized by serum from a patient recently infected by a zoonotic OPV. Taken together, these results demonstrate that chloroplasts are an attractive production vehicle for the expression of OPV subunit vaccines.
| Original language | English |
|---|---|
| Pages (from-to) | 577-591 |
| Number of pages | 15 |
| Journal | Plant Biotechnology Journal |
| Volume | 7 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Aug 2009 |
Fingerprint
Keywords
- A27L
- Chloroplast transformation
- Plant vaccine
- Smallpox vaccine
- Tobacco
- Transgenic plants
ASJC Scopus subject areas
- Plant Science
- Biotechnology
- Agronomy and Crop Science
Cite this
Transgenic chloroplasts are efficient sites for high-yield production of the vaccinia virus envelope protein A27L in plant cells. / Rigano, M. Manuela; Manna, Carmela; Giulini, Anna; Pedrazzini, Emanuela; Capobianchi, Maria; Castilletti, Concetta; Di Caro, Antonino; Ippolito, Giuseppe; Beggio, Paola; De Giuli Morghen, Carlo; Monti, Luigi; Vitale, Alessandro; Cardi, Teodoro.
In: Plant Biotechnology Journal, Vol. 7, No. 6, 08.2009, p. 577-591.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Transgenic chloroplasts are efficient sites for high-yield production of the vaccinia virus envelope protein A27L in plant cells
AU - Rigano, M. Manuela
AU - Manna, Carmela
AU - Giulini, Anna
AU - Pedrazzini, Emanuela
AU - Capobianchi, Maria
AU - Castilletti, Concetta
AU - Di Caro, Antonino
AU - Ippolito, Giuseppe
AU - Beggio, Paola
AU - De Giuli Morghen, Carlo
AU - Monti, Luigi
AU - Vitale, Alessandro
AU - Cardi, Teodoro
PY - 2009/8
Y1 - 2009/8
N2 - Orthopoxviruses (OPVs) have recently received increasing attention because of their potential use in bioterrorism and the occurrence of zoonotic OPV outbreaks, highlighting the need for the development of safe and cost-effective vaccines against smallpox and related viruses. In this respect, the production of subunit protein-based vaccines in transgenic plants is an attractive approach. For this purpose, the A27L immunogenic protein of vaccinia virus was expressed in tobacco using stable transformation of the nuclear or plastid genome. The vaccinia virus protein was expressed in the stroma of transplastomic plants in soluble form and accumulated to about 18% of total soluble protein (equivalent to approximately 1.7 mg/g fresh weight). This level of A27L accumulation was 500-fold higher than that in nuclear transformed plants, and did not decline during leaf development. Transplastomic plants showed a partial reduction in growth and were chlorotic, but reached maturity and set fertile seeds. Analysis by immunofluorescence microscopy indicated altered chlorophyll distribution. Chloroplast-synthesized A27L formed oligomers, suggesting correct folding and quaternary structure, and was recognized by serum from a patient recently infected by a zoonotic OPV. Taken together, these results demonstrate that chloroplasts are an attractive production vehicle for the expression of OPV subunit vaccines.
AB - Orthopoxviruses (OPVs) have recently received increasing attention because of their potential use in bioterrorism and the occurrence of zoonotic OPV outbreaks, highlighting the need for the development of safe and cost-effective vaccines against smallpox and related viruses. In this respect, the production of subunit protein-based vaccines in transgenic plants is an attractive approach. For this purpose, the A27L immunogenic protein of vaccinia virus was expressed in tobacco using stable transformation of the nuclear or plastid genome. The vaccinia virus protein was expressed in the stroma of transplastomic plants in soluble form and accumulated to about 18% of total soluble protein (equivalent to approximately 1.7 mg/g fresh weight). This level of A27L accumulation was 500-fold higher than that in nuclear transformed plants, and did not decline during leaf development. Transplastomic plants showed a partial reduction in growth and were chlorotic, but reached maturity and set fertile seeds. Analysis by immunofluorescence microscopy indicated altered chlorophyll distribution. Chloroplast-synthesized A27L formed oligomers, suggesting correct folding and quaternary structure, and was recognized by serum from a patient recently infected by a zoonotic OPV. Taken together, these results demonstrate that chloroplasts are an attractive production vehicle for the expression of OPV subunit vaccines.
KW - A27L
KW - Chloroplast transformation
KW - Plant vaccine
KW - Smallpox vaccine
KW - Tobacco
KW - Transgenic plants
UR - http://www.scopus.com/inward/record.url?scp=67650308696&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=67650308696&partnerID=8YFLogxK
U2 - 10.1111/j.1467-7652.2009.00425.x
DO - 10.1111/j.1467-7652.2009.00425.x
M3 - Article
C2 - 19508274
AN - SCOPUS:67650308696
VL - 7
SP - 577
EP - 591
JO - Plant Biotechnology Journal
JF - Plant Biotechnology Journal
SN - 1467-7644
IS - 6
ER -