Production and secretion of a heterologous protein by turnip hairy roots with superiority over tobacco hairy roots

A fully contained and efficient heterologous protein production system was designed using Brassica rapa rapa (turnip) hairy roots. Two expression cassettes containing a cauliflower mosaic virus (CaMV) 35S promoter with a duplicated enhancer region, an Arabidopsis thaliana sequence encoding a signal...

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Bibliographic Details
Published in:Biotechnology letters 2014, Vol.36 (1), p.181-190
Main Authors: Huet, Yoann, Ekouna, Jean-Pierre Ele, Caron, Aurore, Mezreb, Katiba, Boitel-Conti, Michèle, Guerineau, François
Format: Article
Language:English
Subjects:
Agricultural biotechnology
Amino Acid Sequence
Applied Microbiology
Arabidopsis thaliana
Base Sequence
Biochemistry
Biomedical and Life Sciences
Biotechnology
Brassica
Brassica napus - genetics
Brassica napus - metabolism
Brassica rapa
Cassettes
Cauliflower mosaic virus
Cauliflowers
Culture
culture media
green fluorescent protein
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Life Sciences
Microbiology
Molecular Sequence Data
Natural products
Nicotiana - genetics
Nicotiana - metabolism
Original Research Paper
Plant Roots - chemistry
Plant Roots - genetics
Plant Roots - metabolism
Plants, Genetically Modified - genetics
Plants, Genetically Modified - metabolism
Protein synthesis
Proteins
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Roots
signal peptide
Studies
Tobacco
Turnips
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Summary:A fully contained and efficient heterologous protein production system was designed using Brassica rapa rapa (turnip) hairy roots. Two expression cassettes containing a cauliflower mosaic virus (CaMV) 35S promoter with a duplicated enhancer region, an Arabidopsis thaliana sequence encoding a signal peptide and the CaMV polyadenylation signal were constructed. One cassette was used to express the green fluorescent protein (GFP)-encoding gene in hairy roots grown in flasks. A stable and fast-growing hairy root line secreted GFP at >120 mg/l culture medium. GFP represented 60 % of the total soluble proteins in the culture medium. Turnip hairy roots retained sustainable growth and stable GFP production over 3 years. These results were superior to those obtained using tobacco hairy roots.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-013-1335-y