A chemically inducible cucumber mosaic virus amplicon system for expression of heterologous proteins in plant tissues.
A novel cucumber mosaic virus inducible viral amplicon (CMViva) expression system has been developed that allows for tightly regulated chemically inducible expression of heterologous genes in plant hosts. Transient production of recombinant alpha(1)-antitrypsin (rAAT), a human blood protein, was demonstrated in Nicotiana benthamiana leaves. The highest production levels were obtained by co-infiltrating leaves with Agrobacterium tumefaciens cells containing CMViva carrying the AAT gene and A. tumefaciens cells carrying a binary vector constitutively expressing the gene silencing suppressor p19. Accumulation of up to thirty-fold more rAAT was observed in leaves (24 mg per 100 g leaf tissue) when compared with the expression levels observed using the cauliflower mosaic virus (CaMV) 35S promoter. Significantly, 70% of the rAAT produced using the CMViva expression system was found to be biologically active, a 170-fold increase in functional protein compared with the CaMV 35S expression system.
Sudarshana
Mysore R
MR
Department of Plant Pathology, University of California at Davis, One Shields Ave., Davis, CA 95616, USA.
Plesha
Michael A
MA
Uratsu
Sandra L
SL
Falk
Bryce W
BW
Dandekar
Abhaya M
AM
Huang
Ting-Kuo
TK
McDonald
Karen A
KA
D016960:Agrobacterium tumefaciens / Q000235:genetics; D017799:Cucumovirus / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D020868:Gene Silencing; D006801:Humans; D010940:Plant Proteins / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D000515:alpha 1-Antitrypsin / Q000235:genetics
D010940:Plant Proteins; D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin