Arabidopsis thaliana hairy roots for the production of heterologous proteins

• Published in: Plant cell, tissue and organ culture Vol. 127; no. 2; pp. 489 - 496
• Main Authors: Mai, Nga T. P., Boitel-Conti, Michèle, Guerineau, François
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01-11-2016; Springer Nature B.V; Springer Verlag
• Subjects: Arabidopsis thaliana; Biomedical and Life Sciences; Elastase; Fluorescence; Genetic resources; Genotype & phenotype; Genotypes; Green fluorescent protein; Hairy root; Hypocotyls; Life Sciences; Original Article; pH effects; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant Sciences; Proteinase inhibitors; Proteins; Roots; Transgenic plants; Yeast; GFP; SERPIN; Protein production; Secretion; Hairy roots
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-016-1073-7

To evaluate the ability of
Arabidopsis thaliana
hairy roots to produce heterologous proteins, hypocotyls were transformed with
Rhizobium rhizogenes
harbouring a green fluorescent protein gene (
gfp
) fused to a plant signal peptide sequence. Hairy root transgenic lines were generated from wild-type or mutant genotypes. A line secreted GFP at 130 mg/l of culture medium. Unlike as was previously found with turnip hairy roots, a His-tag was still attached to approximately 50 % of the protein. Control of the pH and addition of a protease inhibitor to the culture medium resulted in up to 87 % of the GFP retaining the His-tag.
A. thaliana
hairy roots expressing the human serpina1 (α-1-antitrypsin) gene secreted the protein, which was visible on a PAGE gel. Protein activity in the culture medium was demonstrated using an elastase inhibition assay.
A. thaliana
hairy roots can now be considered for the production of heterologous proteins, making it possible to mine the numerous genetic resources for enhancing protein production and quality.