Agrobacterium rhizogenes lacZ-rolC gene expression in Escherichia coli: detection of the product in transgenic plants using RolC-specific antibodies.
The rolC sequence of the Agrobacterium rhizogenes Ri plasmid was fused in-frame to the 3' end of the lacZ gene in plasmid pEX3. The fusion protein RolC-beta-galactosidase was accumulated as insoluble inclusion bodies in Escherichia coli. Antibodies were raised in rabbits against the fusion protein. After affinity purification, RolC-specific antibodies were found to react with a 22-kDa polypeptide prepared from roots of transgenic tobacco plants possessing a rolC gene. The result of differential centrifugation suggested that RolC is present in the soluble fraction of transformed cells.
Department of Botany, Hokkaido University, Sapporo, Japan.
Y
Y
Oono
T
T
Satomi
H
H
Uchimiya
D000595:Amino Acid Sequence; D000906:Antibodies; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics*; D001483:Base Sequence; D002678:Chimera; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis; D012231:Rhizobium / Q000235:genetics*; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D001617:beta-Glucosidase
D000906:Antibodies; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C071257:cytokinin-beta-glucosidase; D001617:beta-Glucosidase; D001616:beta-Galactosidase