Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...
Reexamination Certificate
1999-02-25
2001-01-23
Hutzell, Paula K. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
C435S320100, C435S419000, C435S412000, C435S468000, C536S024100, C800S287000, C800S298000, C800S320000, C800S320100, C800S317000
Reexamination Certificate
active
06177611
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to the field of plant molecular biology, more particularly to regulation of gene expression in plants.
BACKGROUND OF THE INVENTION
Expression of heterologous DNA sequences in a plant host is dependent upon the presence of an operably linked promoter that is functional within the plant host. Choice of the promoter sequence will determine when and where within the organism the heterologous DNA sequence is expressed. Thus, where continuous expression is desired throughout the cells of a plant, constitutive promoters are utilized. In contrast, where gene expression in response to a stimulus is desired, inducible promoters are the regulatory element of choice. In either case, additional regulatory sequences upstream and/or downstream from the core promoter sequence may be included in expression constructs of transformation vectors to bring about varying levels of constitutive or inducible expression of heterologous nucleotide sequences in a transgenic plant.
Frequently it is desirable to have constitutive expression of a DNA sequence throughout the cells of an organism. For example, increased resistance of a plant to infection by soil- and air-borne pathogens might be accomplished by genetic manipulation of the plant's genome to comprise a constitutive promoter operably linked to a heterologous pathogen-resistance gene such that pathogen-resistance proteins are continuously expressed throughout the plant's tissues.
Alternatively, it might be desirable to inhibit expression of a native DNA sequence within a plant's tissues to achieve a desired phenotype. In this case, such inhibition might be accomplished with transformation of the plant to comprise a constitutive promoter operably linked to an antisense nucleotide sequence, such that constitutive expression of the antisense sequence produces an RNA transcript that interferes with translation of the mRNA of the native DNA sequence.
Thus, isolation and characterization of constitutive promoters that can serve as regulatory regions for constitutive expression of heterologous nucleotide sequences of interest are needed for genetic manipulation of plants to exhibit specific phenotypic traits.
SUMMARY OF THE INVENTION
Compositions and methods for regulating expression of heterologous nucleotide sequences in a plant are provided. Compositions are novel nucleotide sequences for constitutive plant promoters, more particularly promoters isolated from maize genes encoding histone H2B, metallothionein, alpha-tubulin 3, elongation factor efla, ribosomal protein rps8, chlorophyll a/b binding protein, and glyceraldehyde-3-phosphate dehydrogenase. A method for constitutively expressing a heterologous nucleotide sequence in a plant using the promoter sequences disclosed herein is provided. The method comprises transforming a plant cell with a transformation vector that comprises a heterologous nucleotide sequence operably linked to one of the plant promoters of the present invention and regenerating a stably transformed plant from the transformed plant cell.
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Kim et al, Plant Mol. Biol., vol. 24, pp. 105-117, 1994.
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Montoliu et al., “A Tandem of &agr;-tubulin Genes Preferentially Expressed inRadicular Tissues fromZea Mays,” Plant Molecular Biology,1989, pp. 1-15, vol. 14, Kluwer Academic Publishers, Belgium.
Joanin et al., “Nucleotide Sequence and Expression of Two cDNA Coding for TwoHistone H2B Variants of Maize,”Plant Molecular Biology,1992, pp. 581-588, vol. 20, Kluwer Academic Publishers, Belgium.
Berberich et al., “Molecular Cloning, Characterization and Expression of an Elongation Factor 1&agr; Gene In Maize,”Plant Molecular Biology,1995, pp. 611-615, vol. 29, Kluwer Academic Publishers, Belgium.
Blast Search of H2B histone vs. SwissProt, GenBank, Dbest, and Patents Databases.
Blast Search of alpha-tubulin 3-18 vs. SwissProt, GenBank, Dbest, and Patents Databases.
Blast Search of efla-11 vs. SwissProt, GenBank, Dbest, and Patents Databases.
Blast Search of efla-15 vs. SwissProt, GenBank, Dbest, and Patents Databases.
Blast Search of efla-16 vs. SwissProt, GenBank, Dbest, and Patents Databases.
Blast Search of rps8 vs. SwissProt, GenBank, Dbest, and Patents Databases.
Blast Search of cab-10 vs. SwissProt, GenBank, Dbest, and Patents Databases.
Blast Search of cab-20 vs. SwissProt, GenBank, Dbest, and Patents Databases.
GenBank Report, Accession No. X69960, Jonin, P., Direct Submission, Submitted Jan. 7, 1993; Joanin, et al., “Molecular Cloning and Sequence Analysis of Two Genes Encoding Two Histone H2B Variants of Maize,” 1994,Physiol. Veg.,vol. 32, pp. 693-696, Sequence from Blast Search for H2Bhistone.
GenBank Report, Accession No. X69961, Joanin, P., Direct Submission, Submitted Jan. 7, 1993; Joanin, et al., “Molecular Cloning and Sequence Analysis of Two Genes Encoding TwoHistone H2B Variants of Maize,” 1994,Physiol. Veg.,vol. 32, pp. 693-696; Sequence from Blast Search for H2Bhistone.
GenBank Report, Accession No. S57628, deFramond, A. J., “A Metallothionein-like Gene from Maize (Zea Mays). Cloning and Characterization,”FEBS Lett.,1991, vol. 290 (1-2), pp. 103-106; Sequence from Blast Search for metallothionein.
GenBank Report, Accession No. X15704; Puigdomenech, P., Direct Submission, Submitted Jun. 29, 1989; Montoliu, et al., “A Tandem of Alpha-tubulin Genes Preferentially Expressed inRadicular Tissues fromZea mays,” Plant Mol. Biol,1990, vol. 14(1), pp. 1-15; Puigdomenech, P., Direct Submission, Submitted Dec. 15, 1994; Sequence from Blast Search for alphatublin 3-18.
GenBank Report, Accession No. X14794; Sullivan, D. T., Direct Submission, Submitted Jul. 13, 1989; Sullivan et al., “Isolation and Characterization of a Maize Chlorophyll a/b Binding Protein Gene that Produces High Levels of mRNA in the Dark,”Mol. Gen. Genet.,1989, vol. 215(3), pp. 431-440; Sequence from Blast Search for cab-10 and cab-20.
GenBank Report, Accession No. X63205 and S39565; Ray, J. A., Direct Submission, Submitted Nov. 13, 1991; Knight et al., “Isolation of a Gene from Maize Encoding a Chlorophyll a/b-binding Protein,”Plant Mol. Biol.,1992, vol. 19 (3), pp. 533-536; relevant for cab-10 and cab-20.
GenBank Report, Accession No. X53398 and S45324;Viret, J. F., Direct Submission, Submitted Jun. 11, 1990; Becker, et al., “The cab-m7 Gene: A Light-inducible, Mesophyll-Specific Gene of Maize,”Plant Mol. Biol.,vol. 20(1), pp. 49-60; relevant for cab-10 and cab-20.
Kyozuka et al., Anaerobic Induction and Tissue-Specific Expression of Maize Adh1 Promoter inTransgenci Rice Plants and Their Progeny, Mol. Gen. Genet, 1991, pp. 40-48, vol. 228, MGG©Springer-Veriag , Japan.
Joanin, Nucleotide Sequence and Expression of Two cDNA Coding for TwoHistone H2B Variants of Maize, Plant Molecular Biology, 1992, pp. 581-588, vol. 20, ©1992Kluwer Academic Publisher, Belgium.
Brignon et al., Constitutive and Cell-Division-Inducible Protein-DNA Interactions in Two MaizeHistone Gene Promoters, The Plant Journal, 1993, pp. 445-457, vol. 4(3),Cedex, France.
Brignon et al., Nuclease Se
Alston & Bird LLP
Hutzell Paula K.
Mehta Ashwin D.
Pioneer Hi-Bred International , Inc.
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