Generation of transgenic plants expressing antibodies to the environmental pollutant microcystin-LR.
We describe the engineering, regeneration, and characterization of transgenic tobacco plants expressing a recombinant antibody specific to microcystin-LR (MC-LR), the environmental toxin pollutant produced by species of cyanobacteria. The antibody was created by a genetic fusion of the antigen-binding regions of the microcystin-specific single-chain antibody, 3A8, with constant regions from the murine IgG1kappa, Guy's 13. IgG transgenes were controlled by a leader peptide that targets the transgene products to the secretory pathway and also allows for rhizosecretion. The antibody, extracted from the leaves or rhizosecreted into hydroponic medium by transgenic plants, was shown to have functional binding to MC-LR. Antibody yields in transgenic plant leaves reached a maximum of 64 microg/g leaf fresh weight (0.6% total soluble protein), and the rate of antibody rhizosecretion reached a maximum of 5 microg/g root dry weight/24 h. Rhizosecreted antibody bound to MC-LR in hydroponic medium, and transgenic plants grew more efficiently on medium containing MC-LR compared to wild-type controls. This proof of concept paves the way for applications to produce diagnostic antibodies to microcystin-LR, remove it from the environment by phytoremediation, or enhance yields in crops exposed to MC-LR.-Drake, P. M. W., Barbi, T., Drever, M. R., van Dolleweerd, C. J., Porter, A. J. R., Ma, J. K.-C. Generation of transgenic plants expressing antibodies to the environmental pollutant microcystin-LR.
Molecular Immunology Unit, Centre for Infection, Department of Cellular and Molecular Medicine, Room 2.148, Floor 2, Jenner Wing, St. George's University of London, Cranmer Terr., London SW17 0RE, UK. pdrake@sgul.ac.uk
Pascal M W
PM
Drake
Tommaso
T
Barbi
Matthew R
MR
Drever
Craig J
CJ
van Dolleweerd
Andrew J R
AJ
Porter
Julian K-C
JK
Ma
D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004785:Environmental Pollutants / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D008387:Marine Toxins; D052998:Microcystins / Q000276:immunology*; D008957:Models, Genetic; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
D000906:Antibodies; D004785:Environmental Pollutants; D008387:Marine Toxins; D052998:Microcystins; D011994:Recombinant Proteins; C057862:cyanoginosin LR