Increasing Expression of an Anti-Picloram Single-Chain Variable Fragment (ScFv) Antibody and Resistance to Picloram in Transgenic Tobacco ( Nicotiana tabacum )
2005; American Chemical Society; Volume: 53; Issue: 17 Linguagem: Inglês
10.1021/jf0507691
ISSN1520-5118
AutoresFernando C. Olea-Popelka, Michael D. McLean, Jonathan Horsman, Kurt C. Almquist, Jim Brandle, J. Christopher Hall,
Tópico(s)CRISPR and Genetic Engineering
ResumoSystematic research involving four chimeric gene constructions designed to express the same anti-picloram single-chain variable fragment (scFv) antibody is described. Agrobacterium-mediated transformation produced at least 25 transgenic tobacco plants with each of these, and the number of T-DNA loci in each plant was determined using kanamycin-resistance segregation assays. The relative amounts of active and total scFv in each plant were evaluated using quantitative enzyme-linked immunosorbent assay and immunoblot technologies, respectively. No significant differences in scFv activity were found among the four groups of single-locus plants, although the 35S/M construct was found to produce significantly more total anti-picloram scFv than the other three constructs. A dose-response bioassay involving T(1) seedlings from several of the highest expressers of active scFv demonstrated resistance to a constant exposure of picloram at 5 x 10(-)(8) M. Other approaches for increasing antibody-based herbicide resistance are discussed, as further improvements are needed before practical application of this technology.
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