High-efficiency expression of a receptor-binding domain of SARS-CoV spike protein in tobacco chloroplasts.
- Author:
Xue ZHONG
;
Guangxun QI
;
Jing YANG
;
Guojie XING
;
Jianfeng LIU
;
Xiangdong YANG
- Publication Type:Journal Article
- MeSH:
Chloroplasts;
metabolism;
Cholera Toxin;
Protein Interaction Domains and Motifs;
Recombinant Fusion Proteins;
biosynthesis;
SARS Virus;
Spike Glycoprotein, Coronavirus;
biosynthesis;
Tobacco;
metabolism;
Vaccines, Subunit;
biosynthesis
- From:
Chinese Journal of Biotechnology
2014;30(6):920-930
- CountryChina
- Language:Chinese
-
Abstract:
Chloroplast-based expression system is promising for the hyper-expression of plant-derived recombinant therapeutic proteins and vaccines. To verify the feasibility of obtaining high-level expression of the SARS subunit vaccine and to provide a suitable plant-derived vaccine production platform against the severe acute respiratory syndrome coronavirus (SARS-CoV), a 193-amino acid fragment of SARS CoV spike protein receptor-binding domain (RBD), fused with the peptide vector cholera toxin B subunit (CTB), was expressed in tobacco chloroplasts. Codon-optimized CTB-RBD sequence was integrated into the chloroplast genome and homoplasmy was obtained, as confirmed by PCR and Southern blot analysis. Western blot showed expression of the recombinant fusion protein mostly in soluble monomeric form. Quantification of the recombinant fusion protein CTB-RBD was conducted by ELISA analysis from the transplastomic leaves at different developmental stages, attachment positions and time points in a day and the different expression levels of the CTB-RBD were observed with the highest expression of 10.2% total soluble protein obtained from mature transplastomic leaves. Taken together, our results demonstrate the feasibility of highly expressing SARS subunit vaccine RBD, indicating its potential in subsequent development of a plant-derived recombinant subunit vaccine and reagents production for antibody detection in SARS serological tests.
