Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...
Reexamination Certificate
1999-12-13
2001-12-18
Fox, David T. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
C435S320100, C435S419000, C435S468000, C536S024100, C800S298000, C800S306000, C800S320000
Reexamination Certificate
active
06331663
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to highly effective modified full-length promoters for plants, such as soybeans and rice.
BACKGROUND OF THE INVENTION
Recent advances in genetic engineering have provided plant breeders and geneticists with the tools to insert or transform genes, which are selected portions of deoxyribonucleic acid (also known as DNA), into a plant in order to produce new kinds of plants known as transgenic plants. Such transgenic plants or crops can have unique characteristics or traits, including resistance to plant diseases, resistance to herbicides, resistance to insects, enhanced stability or shelf-life of the ultimate consumer product obtained from the plant and/or improvements in the nutritional value in the edible portions of the plant. Genes are made up of DNA, a complex molecule inside each plant cell that provides the instructions for all aspects of the plant's growth. A promoter is a region on a gene where transcription factors can bind to enable the gene to “express” itself through the production of another, but smaller molecule known as messenger RNA. Messenger RNA enables the gene to “deliver” its message or instructions to other parts of the plant cell in many cases by being translated into a protein. Various plant promoters have been identified and isolated from different plants, as described in various patents, such as U.S. Pat. Nos. 5,536,653; 5,589,583; 5,608,150; and 5,898,096. Although effective, such promoters have not been modified or optimized to provide enhanced or improved characteristics or traits. It would be desirable to provide plant promoters that have been modified to advantageously provide improved characteristics or traits in plants.
SUMMARY OF THE INVENTION
The present invention relates to a modified promoter that when placed upstream of a gene of interest, will cause that gene to be expressed in plant tissues. The promoter should be active during most of the plant's developmental stages from the seedling stage to maturity.
In a first embodiment, the present invention is directed to a DNA molecule that is SEQ ID NO: 10, wherein nucleotide “w” is “a or t/u” (adenine or thymine/uracil); and nucleotide “y” is “t/u or c”(thymine/uracil or cytosine). Preferably, the DNA molecule is SEQ ID NO: 11 and SEQ ID NO: 20, as follows:
aatgaagtcc caatcctagt ggtcactttt gaaaggaacc ttggcaaagg agggtggctt
[SEQ ID NO:10]
gcaagtgcca tccttgcrat aaaggaaagg tgaaggatca aggtgcccct accaamagtg
gtcttgcctt tggtccccca cccaagagga gcatcctagt acaagtgggg aatccaacca
cgtcttcgca agwggrttgw tgtgayatct ccattgaygt aagggatgac gcacaaagtg
actatccttc acaagacctt cctctatata aggaggggtt cattcccatt tgaaggatca
ata
aatgaagtcc caatcctagt ggtcactttt gaaaggaacc ttggcaaagg agggtggctt
[SEQ ID NO:11]
gcaagtgcca tccttgcgat aaaggaaagg tgaaggatca aggtgcccct accaacagtg
gtcttgcctt tggtccccca cccaagagga gcatcctagt acaagtgggg aatccaacca
cgtcttcgca agagggttgt tgtgacatct ccattgatgt aagggatgac gcacaaagtg
actatccttc acaagacctt cctctatata aggaggggtt cattcccatt tgaaggatca
ata
aatgaagtcc caatcctagt ggtcactttt gaaaggaacc ttggcaaagg agggtggctt
[SEQ ID NO:20]
gcaagtgcca tccttgcaat aaaggaaagg tgaaggatca aggtgcccct accaaaagtg
gtcttgcctt tggtccccca cccaagagga gcatcctagt acaagtgggg aatccaacca
cgtcttcgca agtggattga tgtgatatct ccattgacgt aagggatgac gcacaaagtg
actatccttc acaagacctt cctctatata aggaggggtt cattcccatt tgaaggatca
ata
In its double stranded form, SEQ ID NOS: 10, 11 and 20 are useful as full length promoters in plants, such as dicots and monocots.
In a second embodiment, the present invention is directed toward a DNA construct comprising a DNA molecule that contains SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 20. Preferably the DNA construct is a plasmid. Also preferred is that the plasmid is of the designation pPG345 or pPG346.
In a third embodiment, the present invention is directed toward a eukaryotic cell comprising a DNA molecule that contains SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 20. Preferably, the eukaryotic cell is a plant cell. Also preferred is that the eukaryotic plant cell is a dicot plant cell. However, the eukaryotic plant cell may also be a monocot plant cell.
In a fourth embodiment, the present invention is directed toward a plant having eukaryotic cells comprising a DNA molecule that contains SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 20. Preferably, the plant is a dicot plant, although the plant may also be a monocot plant. Also preferred is that the dicot plant is
Arabidopsis thaliana.
In a fifth embodiment, the present invention is directed toward seed capable of producing a plant having cells comprising a DNA molecule that contains SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 20. Preferably, the seed is capable of producing a plant that is a dicot plant, although the seed may also be able to produce a plant that is a monocot plant.
In a sixth embodiment, the present invention is directed toward a method of controlling and/or increasing the transcription of a heterologous or homologous gene in a plant or plant tissue comprising transforming the plant or plant tissue with a DNA construct comprising a heterologous or homologous gene and a DNA molecule that is SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 20.
One advantage of the present invention is that it provides a promoter that can better resist cytosin methylation (at CG or CNG di- or tri-nucleotides) which can deactivate or reduce the effectiveness of the promoter, compared to other known promoters, such as from double-stranded DNA viruses.
REFERENCES:
patent: 5097025 (1992-03-01), Benfey et al.
patent: 5312910 (1994-05-01), Ganesh et al.
patent: 0426641 (1990-10-01), None
patent: 9800534 (1998-01-01), None
patent: 9927119 (1999-06-01), None
patent: 9943938 (1999-09-01), None
P. Benefey et al, “The Caulflower Mosaic Virus 35S Promoter: Combinatorial Regulatioin of Transcription in Plants”, Nov. 1990, Science vol. 250, pp. 959-966.*
Y. Kim et al, “a 20 nucleotide upstream element is essential for the nopaline synthase (nos) promoter activity”, 1994, Plant Molecular Biology vol. 24, pp 105-117.
Collins Cynthia
Fox David T.
Majka Joseph T.
Nowak Henry P.
Paradigm Genetics, Inc.
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