Rapid transient expression of cholera toxin B subunit (CTB) in Nicotiana benthamiana

Kaewta Rattanapisit, Seong Hee Bhoo, Tae Ryong Hahn, Hugh Mason, Waranyoo Phoolcharoen

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Cholera toxin B subunit (CTB) has the potential to be an effective adjuvant for mucosal vaccines because of its ability to increase antigen uptake and presentation by antigen-presenting cells through GM1-ganglioside binding. CTB has been produced using different recombinant protein expression systems. This study used the geminiviral replicon system to transiently express CTB in Nicotiana benthamiana. The plant-optimized CTB gene was cloned into a geminiviral vector and infiltrated into N. benthamiana leaves. The highest CTB protein level was observed on day 4 with approximately 4 μg/g fresh weight. The Western blot analysis using anti-CTB suggests assembly of CTB into oligomers. Based on the GM1-ELISA results, this CTB transiently expressed in plants showed biological activity for binding the intestinal epithelial cell membrane glycolipid receptor, GM1-glanglioside, which implies its potential as an adjuvant for mucosal vaccines.

Original languageEnglish (US)
Pages (from-to)107-113
Number of pages7
JournalIn Vitro Cellular and Developmental Biology - Plant
Volume49
Issue number2
DOIs
StatePublished - 2013

Fingerprint

cholera toxin
Nicotiana benthamiana
Cholera Toxin
Tobacco
adjuvants
Vaccines
vaccines
G(M1) Ganglioside
gangliosides
replicon
Replicon
phytotoxins
antigen-presenting cells
glycolipids
Antigen Presentation
Antigen-Presenting Cells
protein subunits
Cell membranes
Bioactivity
Recombinant Proteins

Keywords

  • Cholera toxin B subunit (CTB)
  • Geminiviral replicon system
  • Nicotiana benthamiana
  • Transient expression

ASJC Scopus subject areas

  • Plant Science
  • Biotechnology

Cite this

Rapid transient expression of cholera toxin B subunit (CTB) in Nicotiana benthamiana. / Rattanapisit, Kaewta; Bhoo, Seong Hee; Hahn, Tae Ryong; Mason, Hugh; Phoolcharoen, Waranyoo.

In: In Vitro Cellular and Developmental Biology - Plant, Vol. 49, No. 2, 2013, p. 107-113.

Research output: Contribution to journalArticle

Rattanapisit, Kaewta ; Bhoo, Seong Hee ; Hahn, Tae Ryong ; Mason, Hugh ; Phoolcharoen, Waranyoo. / Rapid transient expression of cholera toxin B subunit (CTB) in Nicotiana benthamiana. In: In Vitro Cellular and Developmental Biology - Plant. 2013 ; Vol. 49, No. 2. pp. 107-113.
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