Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part
Patent
1996-01-16
1999-10-05
Fox, David T.
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
Higher plant, seedling, plant seed, or plant part
800278, 800288, 800294, 800298, 435 691, 435193, 4352522, 4353201, 435418, 435419, 435468, 435469, 536 237, H01H 500, C12N 1531, C12N 1554, C12N 1582, A01H 510
Patent
active
059627689
DESCRIPTION:
BRIEF SUMMARY
This invention relates to a chimeric selectable marker gene comprising: a plant-expressible promoter, DNA encoding an aminoglycoside-6'-N-acetyltransferase (the "AAC(6')"), and a 3' end formation and polyadenylation region active in plant cells.
This invention further relates to a process for selecting or identifying transformed plant cells by expressing the chimeric marker gene, encoding the AAC(6'), in the plant cells. The chimeric marker gene confers, on the plant cells, resistance to normally lethal or growth-suppressive concentrations of an antibiotic which is efficiently detoxified by the AAC(6') in the cells.
This invention also relates to a plant cell, stably transformed with the chimeric marker gene, encoding the AAC(6'), and to a plant regenerated from this plant cell.
BACKGROUND OF THE INVENTION
Plant genetic engineering technology has made significant progress during the last decade. It has become possible to introduce stably foreign genes into plants. This has provided exciting opportunities for modern agriculture.
The use of chimeric selectable marker genes in plant cell transformation has considerably simplified the selection of transformed plant cells. For example, by the expression of such a marker gene, transformed plant cells can be made resistant to antibiotics that are cytotoxic or growth-suppressing to non-transformed cells. A commonly used chimeric marker gene contains the neomycin phosphotransferase-II or nptII coding region (Bevan et al (1983) Nature 304, 184-187: Fraley et al (1983) Proc. Natl. Acad. Sci USA 80, 4803-4807). The nptII gene confers resistance to kanamycin, neomycin and G-418 antibiotics on plant cells expressing the gene (Reynaerts et al (1987) Plant Mol. Biol. Manual, Gelvin, S. B. & Schilperoort, R. A. (eds), Kluwer, Dordrecht, sect. A9, pp. 1-16).
Chimeric marker genes have typically contained: a plant-expressible promoter (with a 5' untranslated region); DNA (such as the npt II gene) encoding a selectable marker; and a 3' end formation and polyadenylation region active in plants. Although the versatility of the nptII gene has been confirmed in chimeric marker genes in several plant systems over the years, there have been limitations on its use that have necessitated the development of alternative antibiotic-resistance genes for use in such chimeric selectable marker genes (Hayford et al (1988) Plant Physiol. 86, 1216). Furthermore, in many situations, a second complementary antibiotic-resistance gene has been needed for introduction into plants that have already been transformed with an antibiotic-resistance gene. Such alternative antibiotic-resistance genes already exist, but they often require the use of very toxic substrates and/or they do not allow efficient selection in all plant species. Certainly for species that are routinely vegetatively reproducible, like potato, antibiotic-resistance genes encoding different selectable markers, with different specific substrates, are required when different genes have to be engineered at different times into a plant.
Among the known antibiotic-resistance genes are those encoding aminoglycoside antibiotic-acetylating (AAC) enzymes, four types of which have been characterized (based on the position of the modified amino group of the 2-deoxystreptamine-derived aminoglycosides):AAC(1), AAC(2'), AAC(3) and AAC(6'). See Shaw et al (1989) Antimicrob. Agents & Chemotherapy 33, 2052-2062. High-pressure liquid chromatography (HPLC) analysis has demonstrated the differences among the acetylated products of these four types of enzymes, and aminoglycoside-resistance profiles can be used to identify the presence of each of these types of enzymes in a host strain (Shaw et al (1989) supra).
European patent publication ("EP") 0 289 478 (Rogers et al (1988), Hayford et al (1988) supra, and Carrer et al (1991) Plant Mol. biol. 17, 301-303 describe the selection on gentamycin of plants transformed with an aminoglycoside-3-N-acetyltransferase-encoding gene (the "aac(3) gene"). The aac(3)-IV gene was found to confer resistance to kanamy
REFERENCES:
Bevan et al., Nature, 304, 184-187 (1983).
Fraley et al., Proc. Nat. Acad. Sci. USA, 80, 4803-4807 (1983).
Reynaerts et al., Plant Mol. Biology Manual, PMAN-A9 (1987).
Hayford et al., Plant Physiol. 86, 1216-1222 (1988).
Shaw et al., Antimicrobial Agents and Chemotherapy, 33, 2052-2062 (1989).
Carrer et al., Plant Mol. Biology, 17, 301-303 (1991).
Davies, Antibiotics in Laboratory Medicine, 474-489 (ed. V. Lorian).
Phillips et al., British Medical Bulletin, 40, No. 1, 28-35 (1984).
DeBlock et al., The EMBO Journal, 6, No. 9, 2513-2518 (1987).
Nobuta et al., J. Bacteriology, 170, No. 8, 3769-3773 (1988).
Tolmasky, Plasmid, 24, 218-226 (1990).
Van Vhieu et al., J. Bacteriology, 169, No. 12, 5708-5714 (1987).
Davies et al., Ann Rev. Microbiol., 32, 469-518 (1978).
Morohoshi et al., Journal of Antibioties, XXXVII, No. 12, 1687-1691 (1984).
Tenover et al., J. Bacteriology, 170, No. 1, 471-473 (1988).
Ferretti et al., J. Bacteriology, 167, No. 2, 631-638 (1986).
Leemans et al., Gene, 19, 361-364 (1982).
Cornelissen Marcus
Gossele Veronique
Reynaerts Arlette
Van Aarssen Roel
LandOfFree
Marker gene does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Marker gene, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Marker gene will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1173726