Transient protein expression in three Pisum sativum (green pea) varieties.
The expression of proteins in plants both transiently and via permanently transformed lines has been demonstrated by a number of groups. Transient plant expression systems, due to high expression levels and speed of production, show greater promise for the manufacturing of biopharmaceuticals when compared to permanent transformants. Expression vectors based on a tobacco mosaic virus (TMV) are the most commonly utilized and the primary plant used, Nicotiana benthamiana, has demonstrated the ability to express a wide range of proteins at levels amenable to purification. N. benthamiana has two limitations for its use; one is its relatively slow growth, and the other is its low biomass. To address these limitations we screened a number of legumes for transient protein expression. Using the alfalfa mosaic virus (AMV) and the cucumber mosaic virus (CMV) vectors, delivered via Agrobacterium, we were able to identify three Pisum sativum varieties that demonstrated protein expression transiently. Expression levels of 420 +/- 26.24 mg GFP/kgFW in the green pea variety speckled pea were achieved. We were also able to express three therapeutic proteins indicating promise for this system in the production of biopharmaceuticals.
Green
Brian J
BJ
Fraunhofer USA Center for Molecular Biotechnology, Newark, DE 19711, USA.
Fujiki
Masaaki
M
Mett
Valentina
V
Kaczmarczyk
Jon
J
Shamloul
Moneim
M
Musiychuk
Konstantin
K
Underkoffler
Susan
S
Yusibov
Vidadi
V
Mett
Vadim
V
D017794:Alfalfa mosaic virus / Q000235:genetics*; D017799:Cucumovirus / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D018532:Pisum sativum / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D013045:Species Specificity; D014162:Transfection / Q000379:methods*
D011994:Recombinant Proteins