Comparison of transient protein expression in tobacco leaves and plant suspension culture

Lauren B. Andrews, Wayne R. Curtis

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

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 Nicotiana benthamiana 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.

Original languageEnglish (US)
Pages (from-to)946-952
Number of pages7
JournalBiotechnology Progress
Volume21
Issue number3
DOIs
StatePublished - May 1 2005

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Plant Leaves
Agrobacterium
Nicotiana glutinosa
Tobacco
Suspensions
tobacco
Nicotiana benthamiana
protein synthesis
coculture
cell suspension culture
plant tissues
Cell Culture Techniques
Plant Cells
Coculture Techniques
brushes
auxotrophs
leaves
Proteins
aseptic conditions
Tissue Culture Techniques

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

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abstract = "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 Nicotiana benthamiana 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.",
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Comparison of transient protein expression in tobacco leaves and plant suspension culture. / Andrews, Lauren B.; Curtis, Wayne R.

In: Biotechnology Progress, Vol. 21, No. 3, 01.05.2005, p. 946-952.

Research output: Contribution to journalArticle

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AB - 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 Nicotiana benthamiana 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.

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