Non-food/feed seeds as biofactories for the high-yield production of recombinant pharmaceuticals.
Plant Biotechnol J
; 9(8): 911-21, 2011 Oct.
Article
em En
| MEDLINE
| ID: mdl-21481135
ABSTRACT
We describe an attractive cloning system for the seed-specific expression of recombinant proteins using three non-food/feed crops. A vector designed for direct subcloning by Gateway® recombination was developed and tested in Arabidopsis, tobacco and petunia plants for the production of a chimeric form (GAD67/65) of the 65 kDa isoform of glutamic acid decarboxylase (GAD65). GAD65 is one of the major human autoantigens involved in type 1 diabetes (T1D). The murine anti-inflammatory cytokine interleukin-10 (IL-10) was expressed with the described system in Arabidopsis and tobacco, whereas proinsulin, another T1D major autoantigen, was expressed in Arabidopsis. The cost-effective production of these proteins in plants could allow the development of T1D prevention strategies based on the induction of immunological tolerance. The best yields were achieved in Arabidopsis seeds, where GAD67/65 reached 7.7% of total soluble protein (TSP), the highest levels ever reported for this protein in plants. IL-10 and proinsulin reached 0.70% and 0.007% of TSP, respectively, consistent with levels previously reported in other plants or tissues. This versatile cloning vector could be suitable for the high-throughput evaluation of expression levels and stability of many valuable and difficult to produce proteins.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proinsulina
/
Sementes
/
Vetores Genéticos
/
Glutamato Descarboxilase
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Plant Biotechnol J
Assunto da revista:
BIOTECNOLOGIA
/
BOTANICA
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Itália