RESUMO

Transgenic plants are able to express molecules with antigenic properties. In recent years, this has led the pharmaceutical industry to use plants as alternative systems for the production of recombinant proteins. Plant-produced recombinant proteins can have important applications in therapeutics, such as in the treatment of rheumatoid arthritis (RA). In this study, the mycobacterial HSP65 protein expressed in tobacco plants was found to be effective as a treatment for adjuvant-induced arthritis (AIA). We cloned the hsp65 gene from Mycobacterium leprae into plasmid pCAMBIA 2301 under the control of the double 35S promoter from cauliflower mosaic virus. Agrobacterium tumefaciens bearing the pChsp65 plasmid was used to transform tobacco plants. Incorporation of the hsp65 gene was confirmed by PCR, reverse transcription-PCR, histochemistry, and western blot analyses in several transgenic lines of tobacco plants. Oral treatment of AIA rats with the HSP65 protein allowed them to recover body weight and joint inflammation was reduced. Our results suggest a synergistic effect between the HSP65 expressed protein and metabolites presents in tobacco plants.

Assuntos

Artrite Experimental/tratamento farmacológico , Proteínas de Bactérias/uso terapêutico , Chaperonina 60/uso terapêutico , Nicotiana/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Administração Oral , Agrobacterium tumefaciens/genética , Animais , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Western Blotting , Chaperonina 60/administração & dosagem , Chaperonina 60/genética , Chaperonina 60/metabolismo , Humanos , Mycobacterium leprae/genética , Mycobacterium leprae/metabolismo , Plantas Geneticamente Modificadas/genética , Plasmídeos , Ratos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Nicotiana/genética , Resultado do Tratamento

RESUMO

A highly efficient somatic embryogenesis system and subsequent plant regeneration of chinaberry (Melia azedarach L.) was developed. Plants were regenerated from indirect somatic embryogenesis induction. Novel features of this improved protocol, include: a) Embryogenic callus induction with no addition of 2, 4-D in the culture media; b) Somatic embryos differentiation was achieved by using high concentration of cytokinins (BAP 10 mg/L) and adenine; c) 100 percent conversion of somatic embryos to plants was practically obtained and 100 percent of plants survived under greenhouse conditions; d) Addition of putrescine improved somatic embryos germination. The amount of somatic embryos produced by the pathway of indirect somatic embryogenesis was 447 per gram of fresh weight callus. Regenerated plants were phenotypically normal. The developed protocol established the potential to produce plantlets from cotyledon explants through somatic embryogenesis. It also presents itself as a highly efficient method for mass clonal propagation and conservation of Melia azedarach.