Scale-up of transgenic tobacco cells that express intimin of enterohemorrhagic Escherichia coli O157:H7 for use as a transitional platform for an oral cattle vaccine

Kristin M. O’Neill, Anne M. Schilthuis, Calvin A. Leiter, Kurt M. Neihaus, Nicole A. Judge, Edda Twiddy, Alison D. O’Brien, Wayne R. Curtis

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Intimin is the primary adhesin protein of enterohemorrhagic Escherichia coli O157:H7, a pathogen carried by cattle and transmitted to humans via contaminated food and water. This work describes the rapid production of 1.3 kg dry weight of transgenic tobacco cells expressing the C-terminal 261 amino acids of intimin (Int261) for subsequent oral vaccine testing to reduce cattle colonization by E. coli O157:H7. Both a stirred-tank bioreactor (60 L) and a less capital-intensive oxygenated carboy culture were shown feasible for generating this scale of transgenic plant tissue culture biomass. Antibiotic selection pressure was not required during scale-up to maintain Int261 expression over the 13-wk culture period. Extended medium autoclave times of up to 90 min used for bioreactor sterilization had only minimal impact on nutrient uptake, culture growth, and intimin expression. Plant tissue was transformed, produced in vitro, and available for feeding studies in a fraction of the time required to develop and grow transgenic plants.

Original languageEnglish (US)
Pages (from-to)315-323
Number of pages9
JournalIn Vitro Cellular and Developmental Biology - Plant
Volume51
Issue number3
DOIs
StatePublished - Jun 19 2015

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intimin
Enterohemorrhagic Escherichia coli
enterohemorrhagic Escherichia coli
Escherichia coli O157
Genetically Modified Plants
Bioreactors
Tobacco
mouth
tobacco
Vaccines
genetically modified organisms
vaccines
bioreactors
plant tissues
transgenic plants
cattle
Biomass
autoclaves
adhesins
Economics

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Plant Science

Cite this

O’Neill, Kristin M. ; Schilthuis, Anne M. ; Leiter, Calvin A. ; Neihaus, Kurt M. ; Judge, Nicole A. ; Twiddy, Edda ; O’Brien, Alison D. ; Curtis, Wayne R. / Scale-up of transgenic tobacco cells that express intimin of enterohemorrhagic Escherichia coli O157:H7 for use as a transitional platform for an oral cattle vaccine. In: In Vitro Cellular and Developmental Biology - Plant. 2015 ; Vol. 51, No. 3. pp. 315-323.
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Scale-up of transgenic tobacco cells that express intimin of enterohemorrhagic Escherichia coli O157:H7 for use as a transitional platform for an oral cattle vaccine. / O’Neill, Kristin M.; Schilthuis, Anne M.; Leiter, Calvin A.; Neihaus, Kurt M.; Judge, Nicole A.; Twiddy, Edda; O’Brien, Alison D.; Curtis, Wayne R.

In: In Vitro Cellular and Developmental Biology - Plant, Vol. 51, No. 3, 19.06.2015, p. 315-323.

Research output: Contribution to journalArticle

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AU - O’Neill, Kristin M.

AU - Schilthuis, Anne M.

AU - Leiter, Calvin A.

AU - Neihaus, Kurt M.

AU - Judge, Nicole A.

AU - Twiddy, Edda

AU - O’Brien, Alison D.

AU - Curtis, Wayne R.

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