title: Production of recombinant proteins in clonal root cultures using episomal expression vectors.
abstract: We have developed a fully contained system for expressing recombinant proteins that is based on clonal root cultures and episomal expression vectors. Clonal root lines expressing green fluorescent protein (GFP) or human growth hormone were generated from Nicotiana benthamiana leaves infected with the tobacco mosaic virus-based vector 30B after exposure to Agrobacterium rhizogenes. These lines accumulated GFP at over 50 mg per kg fresh tissue, a level that is comparable with other plant production systems in early stage development. Accumulation of both hGH and GFP in the clonal root lines was sustained over a 3-year period, and in the absence of antibiotic selection. This technology shows promise for commercial production of vaccine antigens and therapeutic proteins in contained facilities.
authors.affiliation: Fraunhofer USA Center for Molecular Biotechnology, 9 Innovation Way, Suite 200, Newark, Delaware 19711, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Skarjinskaia
authors.forename: Jason
authors.initials: J
authors.lastname: Karl
authors.forename: Adriana
authors.initials: A
authors.lastname: Araujo
authors.forename: Karen
authors.initials: K
authors.lastname: Ruby
authors.forename: Shailaja
authors.initials: S
authors.lastname: Rabindran
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Streatfield
authors.forename: Vidadi
authors.initials: V
authors.lastname: Yusibov
mesh_terms: D001709:Biotechnology; D002999:Clone Cells / Q000166:cytology; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis*; D019382:Human Growth Hormone / Q000096:biosynthesis*; D006801:Humans; D018515:Plant Leaves / Q000166:cytology / Q000235:genetics / Q000821:virology; D018517:Plant Roots / Q000166:cytology* / Q000235:genetics; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D012231:Rhizobium; D014026:Nicotiana / Q000166:cytology / Q000235:genetics / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins; D019382:Human Growth Hormone; D049452:Green Fluorescent Proteins