Highly efficient expression of Escherichia coli heat-labile enterotoxin B subunit in plants using potato virus X-based vector
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A synthetic gene of the B-subunit of Escherichia coli heat-labile toxin, optimized for expression in plants, was designed and synthesized. The recombinant viral vector was constructed on the basis of potato virus X containing the LTB gene instead of the removed triple block of transport genes and the coat protein gene, which provides for LTB expression in plants. The vector is introduced into the plant cells during cell infiltration by agrobacteria incorporating a binary vector, the T-DNA region of which contains a cDNA copy of the recombinant viral genome. Under conditions of posttranscriptional gene silencing inhibition, the LTB yield in Nicotiana benthamiana plants is 1–2% of total soluble protein; in this case, LTB synthesized in plants forms pentameric complexes analogous to those found in the native toxin. The designed viral system of LTB transient expression can be used to obtain in plants a vaccine against enteropathogenic Escherichia coli.
Key wordsplant as “biofactories” LTB vaccine viral vector
green fluorescent protein
heat-labile toxin of Escherichia coli
B-subunit of Escherichia coli heat-labile toxin
pectin methyl esterase
post-transcriptional gene silencing
potato virus X
RNA-dependent RNA polymerase
35S RNA of cauliflower mosaic virus
synthetic gene of LTB
subgenomic RNA promoter
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- 26.Hirst, T. R. (1995) in Handbook of Natural Toxins, Vol. 8, Bacterial Toxins and Virulence Factors in Disease (Moss, J., Ihlewski, B., Vaughan, M., and Tu, A. T., eds.) Marcel Dekker, N. Y., pp. 123–184.Google Scholar