Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...
Reexamination Certificate
1998-11-12
2003-04-01
Fox, David T. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
C800S287000, C800S288000, C800S317000, C800S317200, C800S323100, C435S417000, C435S419000, C435S429000, C435S430000, C435S468000
Reexamination Certificate
active
06541682
ABSTRACT:
TECHNICAL FIELD
The invention relates to methods of genetically transforming plant plastids, and more specifically to genetically transforming the plastid genomes of Solanaceous plant species.
BACKGROUND
The plastids of higher plants are an attractive target for genetic engineering. Plant plastids (chloroplasts, amyloplasts, elaioplasts, etioplasts, chromoplasts, etc.) are the major biosynthetic centers that, in addition to photosynthesis, are responsible for production of industrially important compounds such as amino acids, complex carbohydrates, fatty acids, and pigments. Plastids are derived from a common precursor known as a proplastid and thus the plastids present in a given plant species all have the same genetic content. In general, plant cells contain 500-10,000 copies of a small 120-160 kilobase circular genome, each molecule of which has a large (approximately 25 kb) inverted repeat. Thus, it is possible to engineer plant cells to contain up to 20,000 copies of a particular gene of interest which potentially can result in very high levels of foreign gene expression. In addition, plastids of most plants are maternally inherited. Consequently, unlike heterologous genes expressed in the nucleus, heterologous genes expressed in plastids are not pollen disseminated, therefore, a trait introduced into a plant plastid will not be transmitted to wild-type relatives.
Unfortunately, successful chloroplast transformation techniques described thusfar have been limited to tobacco (U.S. Pat. No.5,451,513; Svab et. al. (1990),
Proc. Natl. Acad. Sci. USA
87:8526-8530 and Svab et al. (1993),
Proc. Natl. Acad. Sci. USA
90:913-197). For practical applications of genetic engineering techniques to crop plant plastids, chloroplast transformation techniques for a wide variety of crop plants are needed in the art.
The genus Solanaceae includes many agriculturally important plants, and includes some 95 genera. Solanaceous lid crop plants include potato, tomato, eggplant, and other lesser known edible fruits from Physalis (cape gooseberry, strawberry tomato, jamberberry, sugar cherry, chinese lantern, etc), tamarillo, and Capsicum (sweet and chili peppers). The genus also includes many cultivated ornamentals, for example, Petunia, Lycium, Solanum, and Solandra. Other important crops from the genus Solanaceae include tobacco (Nicotiana) and other poisonous alkaloid producing plants such as Hyoscyamus and Datura.
Plastids of higher plants present an attractive target for genetic engineering. As mentioned above, plastids of higher plants are maternally inherited. This offers an advantage for genetic engineering of plants for tolerance or resistance to natural or chemical conditions, such as herbicide tolerance, as these traits will not be transmitted to wild-type relatives.
The production of chloroplast transformation methods applicable to crop species other than tobacco is needed in the art. Such methods provide for a novel means of genetic engineering via plastid transformation to an attractive alternative to nuclear expression of agronomically as well as qualitatively important traits via genetic engineering of plant plastids.
Relevant Literature
Stable transformation of plastids has been reported in the green algae Chlamydomonas (Boynton et al. (1988)
Science
240:1534-1538) and most recently in higher plants (Svab et al. (1990)
Proc. Natl. Acad. Sci. USA
87:8526-8530; Svab and Maliga (1993)
Proc. Natl. Acad. Sci. USA
90:913-917). These methods rely on particle gun delivery of DNA containing a selectable marker and targeting to the plastid genome by homologous recombination.
The complete DNA sequences of the plastid genomes from liverwort (Ohyama et al. (1986)
Nature
322:572-574), rice (Hiratsuka et al. (1989)
Mol. Gen. Genet
. 217:185-194), and tobacco (Shinozaki et al. (1986)
EMBO J
. 5:2043-2049) have been reported.
SUMMARY OF THE INVENTION
The present invention provides methods for the transformation and regeneration of plants containing plant cells, the plastids of which have been stably transformed by a foreign DNA of interest. The method generally comprises transforming a Solanaceous plant cell plastid with a DNA construct; selecting for cells which contain the DNA construct; and obtaining a mature multicellular plant, the cells of which contain the DNA construct in the plant cell plastid.
The instant invention also provides methods for transforming the plastids of Solanaceous plant cells with a DNA construct generally comprising, in the 5′ to 3′ direction of transcription, a promoter region functional in a plant cell plastid, a DNA sequence of interest, and a transcription termination region functional in a plant cell plastid.
Furthermore, the present invention also provides the multicellular solanaceous plant obtained by the methods described herein.
The invention also provides a multicellular solanaceous plant, the plastids of which have been transformed with a DNA construct of interest.
The invention also provides a method for obtaining a plant cell, of which the plastid has been stably transformed with a DNA construct, comprising in the 5′ to 3′ direction of transcription, a promoter functional in a plant cell plastid, a DNA sequence encoding a green fluorescent protein (herein referred to as GFP), and a transcriptional termination region functional in a plant cell plastid.
The invention also provides for the multicellular plant, the plastids of which have been transformed with a DNA construct comprising in the 5′ to 3′ direction of transcription, a promoter functional in a plant cell plastid, a DNA sequence encoding a green fluorescent protein (herein referred to as GFP), and a transcriptional termination region functional in a plant cell plastid.
REFERENCES:
patent: 5015580 (1991-05-01), Christou et al.
patent: 5094945 (1992-03-01), Comai
patent: 5349123 (1994-09-01), Shewmaker et al.
patent: 5349125 (1994-09-01), Holton et al.
patent: 5378619 (1995-01-01), Rogers
patent: 5416011 (1995-05-01), Hinchee et al.
patent: 5451513 (1995-09-01), Maliga et al.
patent: 5545818 (1996-08-01), McBride
patent: 5576198 (1996-11-01), McBride et al.
patent: 5627061 (1997-05-01), Barry et al.
patent: 5633435 (1997-05-01), Barry et al.
patent: 5693507 (1997-12-01), Daniell et al.
patent: 2183660 (1987-06-01), None
patent: WO 92/15675 (1992-09-01), None
patent: WO 99/10513 (1999-03-01), None
Dix et al. Euphytica 85(1-3):29-34, 1995.*
Sidorov et al. Theor. Appl. Genet. 88(3-4): 525-529, 1994.*
Van Grinsven et al. pp. 859-866 In: Genetic Manip. Plant Breed., Horn, W., ed., 1986.*
Perl et al. Plant Mol. Biol. 19:815-823 (No. 5), 1992.*
Chasan, R. Plant Cell 4(1):1-2, Jan. 1992.*
McBride et al. Amplifications of a chimeric bacillus gene in chloroplasts leads to an extraordinary level of an insecticidal protein in tobacco.Biotechnology13:362-365 (1995).
Sheen et al. Green-fluorescent protein as a new vital marker in plant cells.The Plant Journal8(5):777-784 (1995).
Sidorov et al. Stable chloroplast transformation in potato: use of green fluorescent protein as a plastid marker.The Plant Journal19(2):209-216 (1999).
Staub et al. High-yield production of a human therapeutic protein in tobacco chloroplasts.Nature Biotechnology18:333-338 (2000).
Horsch, et al., “A simple and general method for transferring genes into plants”Science227, 1229-1231 (1985).
Klein, et al., “Transformation of microbes, plants and animals by particle bombardment”Bio/Technology10, 286-291 (1992).
Padgette, et al., “Bacterial expression and isolation ofPetunia hybrida5-enol-pyruvylshikimate-3-phosphate synthase”Archives of Biochemistry and Biophysics258(2), 564-573 (1987).
Stalker, et al., “A single amino acid substitution in the enzyme 5-enolpyruvylshikimate-3-phosphate synthase confers resistance to the herbicide glyphosate”The Journal of Biological Chemistry260(8), 4724-4728 (1985).
Stalker, et al., “Purification and properties of a nitrilase specific for the herbicide bromoxynil and corresponding nucleotide sequence analysis of the bxn gene”The Journal of Biological Chemistry2
Nehra Narender S.
Schaaf David J.
Sidorov Vladimir
Stalker David M.
Ye Guangning
Calgene LLC
Fox David T.
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