Chemistry: molecular biology and microbiology – Plant cell or cell line – per se ; composition thereof;... – Plant cell or cell line – per se – contains exogenous or...
Reexamination Certificate
2001-05-24
2003-10-28
Mehta, Ashwin (Department: 1638)
Chemistry: molecular biology and microbiology
Plant cell or cell line, per se ; composition thereof;...
Plant cell or cell line, per se, contains exogenous or...
C536S024100
Reexamination Certificate
active
06638766
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to nucleic acid sequences isolated from sugarcane and to methods of using them. In particular, the inventions relates to nucleotide sequences which are derived from sugarcane polyubiquitin genes and which are capable of directing constitutive expression of a nucleic acid sequence of interest that is operably linked to the sugarcane polyubiquitin nucleotide sequences. The sugarcane polyubiquitin nucleotide sequences are useful in regulating expression of a nucleic acid sequence of interest in monocotyledonous and dicotyledonous plants.
BACKGROUND OF THE INVENTION
Much scientific effort has been directed at genetically engineering plants to produce agronomically relevant proteins. Recombinant genes for producing proteins in plants require a promoter sequence which is capable of directing protein expression in plant cells. Promoter sequences which direct high levels of protein expression in plant cells are particularly desirable since fewer numbers of transgenic plants need to be produced and screened to recover plants producing agronomically significant quantities of the target protein. In addition, high levels of protein expression aid the generation of plants which exhibit commercially important phenotypic properties, such as pest and disease resistance, resistance to environmental stress (e.g., water-logging, drought, heat, cold, light-intensity, day-length, chemicals, etc.), improved qualities (e.g., high yield of fruit, extended shelf-life, uniform fruit shape and color, higher sugar content, higher vitamins C and A content, lower acidity, etc.).
Some promoter sequences which are capable of driving expression of transgenes (e.g. selectable marker genes) in plants are known in the art and are derived from a variety of sources such as bacteria, plant DNA viruses, and plants. Promoters of bacterial origin include the octopine synthase promoter, the nopaline synthase promoter and other promoters derived from native Ti plasmids. Promoters of viral origin include the 35S and 19S RNA promoters of cauliflower mosaic virus. Plant promoters include the ribulose-1,3-diphosphate carboxylase small subunit promoter, the phaseolin promoter, and the maize polyubiquitin promoter.
While some promoter sequences which function in plant cells are available, expression of more than one gene (e.g., a selectable marker gene and an agronomically relevant gene) which is operably linked to the same promoter sequence is likely to be hampered by homology dependent silencing of transgenes in plants. Thus, what is needed are additional promoter sequences which are capable of driving transgene expression. In particular, what is needed are promoter sequences which drive transgene expression in both monocotyledonous and dicotyledonous plant cells.
SUMMARY OF THE INVENTION
The present invention provides nucleic acid sequences having promoter activity. The nucleic acid sequences provided herein direct expression of operably linked nucleotide sequences in cells, tissues and organs of monocotyledonous and dicotyledonous plants. In one embodiment, the invention provides a substantially purified nucleic acid sequence comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:7, the complement of SEQ ID NO:7, homologs of SEQ ID NO:7, homologs of the complement of SEQ ID NO:7; SEQ ID NO:10, the complement of SEQ ID NO:10, homologs of SEQ ID NO:10, and homologs of the complement of SEQ ID NO:10. In a preferred embodiment, the nucleotide sequence is characterized by having promoter activity. In a more preferred embodiment, the promoter activity is constitutive. In another embodiment, nucleotide sequence is double-stranded. In yet another embodiment, the nucleotide sequence is single-stranded. In yet another alternative embodiment, the nucleic acid sequence is contained in a plant cell. In a preferred embodiment, the plant cell is derived from a monocotyledonous plant. In a more preferred embodiment, the monocotyledonous plant is selected from the group consisting of sugarcane, maize, sorghum, pineapple, rice, barley, oat, wheat, rye, yam, onion, banana, coconut, date, and hop. In an alternative more preferred embodiment, the plant cell is derived from a dicotyledonous plant. In a preferred embodiment, the dicotyledonous plant is selected from the group consisting of tobacco, tomato, soybean, and papaya.
The invention also provides a substantially purified nucleic acid sequence comprising a portion of a nucleotide sequence selected from the group consisting of SEQ ID NO:7 and the complement thereof. In a preferred embodiment, the portion is characterized by having promoter activity. In a more preferred embodiment, the promoter activity is constitutive. In an alternative preferred embodiment, the portion is double-stranded. In another alternative preferred embodiment, the portion is single-stranded. In yet another alternative preferred embodiment, the portion comprises the nucleotide sequence selected from the group consisting of the nucleotides from 1 to 242, from 245 to 787, from 788 to 1020, from 1021 to 1084, from 1085 to 1168, from 1169 to 1173, from 1174 to 1648, from 1649 to 1802, from 1 to 377, from 378 to 442, and from 443 to 1802. In a further alternative preferred embodiment, nucleic acid sequence is contained in a plant cell. In a more preferred embodiment, the plant cell is derived from a monocotyledonous plant. In a yet more preferred embodiment, the monocotyledonous plant is selected from the group consisting of sugarcane, maize, sorghum, pineapple, rice, barley, oat, wheat, rye, yam, onion, banana, coconut, date, and hop. In another more preferred embodiment, the plant cell is derived from a dicotyledonous plant. In a yet more preferred embodiment, the dicotyledonous plant is selected from the group consisting of tobacco, tomato, soybean, and papaya.
Further provided by the invention is a substantially purified nucleic acid sequence comprising a portion of a nucleotide sequence selected from the group consisting of SEQ ID NO:10 and the complement thereof. In a preferred embodiment, the portion is characterized by having promoter activity. In a more preferred embodiment, the promoter activity is constitutive. In an alternative preferred embodiment, the portion is double-stranded. In another alternative preferred embodiment, the portion is single-stranded. In yet another alternative preferred embodiment, the portion comprises the nucleotide sequence selected from the group consisting of the nucleotides 1 to 3600, from 3602 to 3612, from 3614 to 3688, from 1 to 2248, from 2249 to 2313, from 2314 to 3688, and from 1671 to 2248. In another alternative preferred embodiment, the nucleic acid sequence is contained in a plant cell. In a more preferred embodiment, the plant cell is derived from a monocotyledonous plant. In yet a more preferred embodiment, the monocotyledonous plant is selected from the group consisting of sugarcane, maize, sorghum, pineapple, rice, barley, oat, wheat, rye, yam, onion, banana, coconut, date, and hop. In an alternative more preferred embodiment, the plant cell is derived from a dicotyledonous plant. In a yet more preferred embodiment, the dicotyledonous plant is selected from the group consisting of tobacco, tomato, soybean, and papaya.
The invention additionally provides a substantially purified nucleic acid sequence comprising the EcoRI/XbaI fragment isolated from plasmid pubi4-GUS contained in
Escherichia coli
cells deposited as NRRLB-30115, the complement of the fragment, homologs of the fragment, and homologs of the complement of the fragment. In a preferred embodiment, the nucleotide sequence is SEQ ID NO:7. In a more preferred embodiment, the nucleotide sequence is characterized by having promoter activity. In a yet more preferred embodiment, the promoter activity is constitutive. In an alternative yet more preferred embodiment, the nucleotide sequence is double-stranded. In another alternative more preferred embodiment, the nucleotide sequence is single-stranded. In yet another alternative more preferred
Albert Henrik H.
Wei Hairong
Connor Margaret A.
Medlen & Carroll LLP
Mehta Ashwin
The United States of America as represented by the Secretary of
LandOfFree
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