Stable and efficient expression of human brain-derived neurotrophic factor in tobacco chloroplasts.

Abstract

Brain-derived neurotrophic factor (BDNF) is an intensively studied neurotrophin that promotes various physiological processes, such as acceleration of cell proliferation and differentiation, and is, therefore widely used in clinical applications. In this study, an expression vector with a codon-optimized hBDNF gene was constructed and transferred into chloroplasts of tobacco by gene-gun. After three or four rounds of selection with optimal spectinomycin concentration, hBDNF was integrated into the chloroplast genome of homoplastomic plants, as confirmed by PCR and Southern hybridization. ELISA indicated that hBDNF fused with GFP represented approximately 15.72% ± 0.33% of total soluble protein in the leaves of transplastomic plants. Moreover, the chloroplast-derived hBDNF displayed biological activity similar to the commercial product. This is the first case report of hBDNF expression by chloroplast transformation in the plant model, providing an additional pathway for the production of chloroplast-expressed therapeutic proteins.