Highly efficient expression of Escherichia coli heat-labile enterotoxin B subunit in plants using potato virus X-based vector.
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.
Ravin
N V
NV
Center Bioengineering, Russian Academy of Sciences, Moscow, 117312, Russia. nravin@biengi.ac.ru
Kuprianov
V V
VV
Zamchuk
L A
LA
Kochetov
A V
AV
Dorokhov
Yu L
YL
Atabekov
J G
JG
Skryabin
K G
KG
D000595:Amino Acid Sequence; D001427:Bacterial Toxins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004768:Enterotoxins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D017863:Potexvirus / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012231:Rhizobium / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014026:Nicotiana / Q000378:metabolism
D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; C027538:heat-labile enterotoxin, E coli