Generation of transgenic plants expressing plasma membrane-bound antibodies to the environmental pollutant microcystin-LR.
In this paper we describe the engineering and regeneration of transgenic tobacco plants expressing a recombinant plasma membrane-retained antibody specific to microcystin-LR (MC-LR), the environmental toxin pollutant produced by cyanobacteria. The antibody was created by a genetic fusion of the antigen binding regions of the microcystin-specific single chain antibody, 3A8, with the constant regions from the murine IgG1κ, Guy's 13, including a membrane retention sequence at the C-terminal end of the antibody heavy chain. The antibody produced in the leaves was shown to be functional by binding to MC-LR in an ELISA with antibody yields in transgenic plant leaves reaching a maximum of 1.2 μg g(-1) leaf f.wt (0.005% total soluble protein). Antibody-MC-LR complexes formed in leaves after addition of MC-LR to hydroponic medium around the roots of transgenic plant cultures.
Barbi
Tommaso
T
Molecular Immunology Unit, Centre for Infection, Division of Clinical Sciences, St. George's University of London, Cranmer Terrace, room 2.148 Floor 2 Jenner Wing, London, SW17 0RE, UK.
Drake
Pascal M W
PM
Drever
Matthew
M
van Dolleweerd
Craig J
CJ
Porter
Andrew R
AR
Ma
Julian K-C
JK
D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism*; D004785:Environmental Pollutants / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D008387:Marine Toxins; D052998:Microcystins / Q000276:immunology*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; 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