Expression of a cholera toxin B subunit in transgenic lettuce (Lactuca sativa L.) using Agrobacterium-mediated transformation system

To increase expression level of cholera toxin B subunit (CTB) in lettuce plants, synthetic CTB (sCTB) gene based on the optimized codon usage was fused with an endoplasmic reticulum retention signal, KDEL. The sCTB gene was introduced into a plant expression vector and transformed to lettuce plants...

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Published in:Plant cell, tissue and organ culture tissue and organ culture, 2006-11, Vol.87 (2), p.203-210
Main Authors: KIM, Young-Sook, KIM, Bang-Genl, KIM, Tae-Geum, KANG, Tae-Jin, YANG, Moon-Sik
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Eukaryotic cell cultures
Fundamental and applied biological sciences. Psychology
Lactuca sativa
Methods. Procedures. Technologies
Miscellaneous
Plant cells and fungal cells
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Summary:To increase expression level of cholera toxin B subunit (CTB) in lettuce plants, synthetic CTB (sCTB) gene based on the optimized codon usage was fused with an endoplasmic reticulum retention signal, KDEL. The sCTB gene was introduced into a plant expression vector and transformed to lettuce plants using Agrobacterium-mediated transformation system. As a selection marker, a bialaphos resistance (bar) gene that encodes phosphinothricin acetyltransferase (PAT), conferring tolerance to the herbicide phosphinothricin (PPT), was used. PCR amplification of genomic DNA confirmed the presence of the sCTB gene in the transgenic lettuce plants. Expressions of mRNA and protein of sCTB were observed by Northern and Western blot analyses, respectively. The sCTB synthesized in the transgenic lettuce showed strong affinity for GM sub(1)-ganglioside suggesting that the sCTB conserved the antigenic sites for binding and proper folding of pentameric CTB structure. The expression level of CTB was relatively high, reaching total soluble protein (TSP) levels of 0.24% in transgenic lettuce.
ISSN:0167-6857
1573-5044
DOI:10.1007/s11240-006-9156-5