Means for identifying a novel class of genes resistant to...

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Separation or purification

Reexamination Certificate

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Reexamination Certificate

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07132514

ABSTRACT:
The invention concerns a method for capturing the target proteins indispensable to the infectious cycles of a pathogenic virus, in particular the rice yellow mottle virus (RYMV) and for cloning the genes involved in said processes. The invention therefore concerns a method for identifying molecular markers of the resistance to RYMV. The method involves the isolation of said protein complexes with viral particles. The method consists in subjecting the samples containing said complexes to electrophoresis and Western Blot using a capsid anti-protein monoclonal antibody, and in recuperating the non-immunodetected bands. The invention also concerns a cDNA capable of being hybridized with a BAC (Bacterial Artificial Chromosome) screened from a bank consisting of DNA fragments of a variety of rice such as IR64. Said BAC clone contains DNA sequences of the markers identified from the rice by means of a process which consists in comparing the AFLP (Amplified Length Polymorphism) of resistant and sensitive rice plants.

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Opalka et al. “Movement of rice yellow mottle virus between xylem cells through pit membranes” Proc. Natl. Acad. Sci. Mar. 1998 vol. 95 pp. 3323-3328.
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Citovsky, V. Tobacco mosaic virus: a pioneer of cell-to-cell movement. Philosophical transactions of the Royal Society of London. Series B, Biological sciences. Mar. 29, 1999, 354 (1383) p. 637-43.
Dorokhov et al. A novel function for a ubiquitous plant enzyme pectin methylesterase: the host-cell receptor for the tobacco mosaic virus movement protein. FEBS letters, Nov. 19, 1999, 461 (3) p. 223-8.
Jackson, D. Opening up the communication channels: Recent insights into plasmodesmal function. Current Opinion in Plant Biology 3 (5): p. 394-399 Oct. 2000.
Bonneau et al, “Expression of the rice yellow mottle virus P1 protein in vitro and in vivo and its Involvement in virus spread”, VIROLOGY, vol. 244, No. 1, Apr. 25, 1998, pp. 79-86.
Opalka et al, “Movement of rice yellow mottle virus between xylem cells through pit membranes”, Proceedings of the National Academy of Sciences of the United States, vol. 95, No. 6, Mar. 17, 1998, pp. 3323-3328.
Albar et al, “Genetic basis and mapping of the resistance to rice yellow mottle virus: I. QTLs identification and relationship between resistance and plant morphology”, Theoretical and Applied Genetics, (Nov. 1998) vol. 97, No. 7, pp. 1145-1154.
Pressoir et al, “Genetic basis and mapping of the resistance to rice yellow mottle virus: II. Evidence of a complementary epistasis between two QTLs”, Theoretical and Applied Genetics, (Nov. 1998) vol. 97, No. 7, pp. 1155-1161.
DATABASE: Wing et al, “A BAC sequencing framework to sequence the rice genome”, Jun. 10, 2000, DATABASE entry/Accession No. AZ132900.

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