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Reduced virus infectivity inN. tabacum secreting a TMV-specific full-size antibody
Authors:Andreas Voss  Marion Niersbach  Rüdiger Hain  Heinz Josef Hirsch  Yu Cai Liao  Fritz Kreuzaler  Rainer Fischer
Institution:(1) Institut für Biologie I, RWTH Aachen, Worringer Weg 1, D-52074 Aachen, Germany;(2) PF-F/Biotechnologie, Bayer AG, D-51368 Leverkusen, Germany
Abstract:We developed a new concept of controlling plant virus infection based on the expression and secretion of full-size antibodies in plants. The neotope-specific anti-Tobacco Mosaic Virus (anti-TMV) antibody mAb24 has a high affinity towards epitopes present only on the surface of intact tobacco mosaic virions. The infectivity of the virus is inhibited almost completely if TMV is adsorbedin vitro at ratios as low as 300 antibody molecules per virion prior to inoculation. Cloned full-size cDNAs of mAb24 heavy and light chains were integrated into the plant expression vector pSS in tandem array and used for transformation ofNicotiana tabacum. The resulting transgenic tobacco plants expressed heavy- and light-chains of mAb24 which were assembled into functional antibodies and exported to the intercellular space. TMV specificity and affinity of the plant-produced antibody (mAb24-P) was not altered when compared to the original murine mAb24. F1 progenies segregated 3:1 with respect to antibody secretion and showed up to two-fold higher expression levels compared to the F0 plants. Upon infection with TMV F1 plants producing mAb24-P showed a reduction of necrotic lesion numbers which is correlated with the amount of antibody produced in transgenic plants.The nucleotide sequence data reported appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession numbers X67210 (heavy-chain cDNA) and X67211 (light-chain cDNA).
Keywords:virus resistance  tobamovirus  recombinant antibodies  transgenic plants  tandem gene expression  35s promoter
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