TY - JOUR
T1 - Rapid transient expression of cholera toxin B subunit (CTB) in Nicotiana benthamiana
AU - Rattanapisit, Kaewta
AU - Bhoo, Seong Hee
AU - Hahn, Tae Ryong
AU - Mason, Hugh
AU - Phoolcharoen, Waranyoo
N1 - Funding Information:
Acknowledgments This work was supported in part by Integrated Innovation Academic Center (IIAC) Chulalongkorn University Center-nary Academic Development Project and the National Research Foundation of Korea (NRF No. 2011-0021436) grant funded to Seong Hee Bhoo by the Korea government.
PY - 2013/4
Y1 - 2013/4
N2 - 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.
AB - 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.
KW - Cholera toxin B subunit (CTB)
KW - Geminiviral replicon system
KW - Nicotiana benthamiana
KW - Transient expression
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U2 - 10.1007/s11627-012-9484-6
DO - 10.1007/s11627-012-9484-6
M3 - Article
AN - SCOPUS:84875412945
SN - 1054-5476
VL - 49
SP - 107
EP - 113
JO - In Vitro Cellular and Developmental Biology - Plant
JF - In Vitro Cellular and Developmental Biology - Plant
IS - 2
ER -