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

Biotechnol Lett. 2014 Jan;36(1):181-90. doi: 10.1007/s10529-013-1335-y.

Abstract

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.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Brassica napus / genetics
  • Brassica napus / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Molecular Sequence Data
  • Nicotiana / genetics
  • Nicotiana / metabolism*
  • Plant Roots / chemistry
  • Plant Roots / genetics
  • Plant Roots / metabolism*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*

Substances

  • Recombinant Proteins
  • Green Fluorescent Proteins