Comparison of transient protein expression in tobacco leaves and plant suspension culture.
Transient gene expression is being developed to provide a more rapid means of assessing plant tissues as a protein production platform without the labor-intensive and time-consuming process of generating stably transformed transgenic plants. Transient expression of the gus-intron reporter gene was facilitated in three different tobacco species. Two different approaches to T-DNA delivery were compared: (1) infiltration of a prototrophic strain of Agrobacterium into leaves and (2) coculture of plant cell suspension cultures with an Agrobacterium auxotroph. Wounding of plant tissues with a wire brush prior to infiltration had a large positive impact on Nicotianabenthamiana leaves but not for Nicotiana tabacum or Nicotiana glutinosa. The best expression level achieved by leaf infiltration was in N. benthamiana (0.025% total soluble protein). A cell suspension culture line of N. glutinosa achieved an expression level greater than 0.04% TSP. The tissue culture-based technique therefore provides improved levels of transient expression under aseptic conditions to facilitate improvements in expression by control of the plant cell culture and Agrobacterium coculture environments.
Andrews
Lauren B
LB
Department of Chemical Engineering, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
Curtis
Wayne R
WR
D002478:Cells, Cultured; D004269:DNA, Bacterial / Q000008:administration & dosage / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D018014:Gene Transfer Techniques; D018515:Plant Leaves / Q000235:genetics* / Q000378:metabolism*; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012231:Rhizobium / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*
D004269:DNA, Bacterial; D010940:Plant Proteins; D011994:Recombinant Proteins; C036483:T-DNA