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AssigneeIowa State University Research Foundation, Inc.
InventorsBASIL J. NIKOLAU, EVE S. WURTELE, DAVID J. OLIVER, ROBERT BEHAL, PATRICK S. SCHNABLE, JINSHAN KE, JERRY L. JOHNSON, CAROLYN C. ALLRED, BETH FATLAND, ISABELLE LUTZIGER, TSUI-JUNG WEN
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The present invention provides nucleic acid and amino acid sequences of acetyl CoA synthetase (ACS), plastidic pyruvate dehydrogenase (pPDH), ATP citrate lyase (ACL), Arabidopsis pyruvate decarboxylase (PDC), and Arabidopsis aldehyde dehydrogenase (ALDH), specifically ALDH-2 and ALDH-4. The present invention also provides a recombinant vector comprising a nucleic acid sequence encoding one of the aforementioned enzymes, an antisense sequence thereto or a ribozyme therefor, a cell transformed with such a vector, antibodies to the enzymes, a plant cell, a plant tissue, a plant organ or a plant in which the level of an enzyme has been altered, and a method of producing such a plant cell, plant tissue, plant organ or plant. Desirably, alteration of the level of enzyme results in an alteration of the level of acetyl CoA in the plant cell, plant tissue, plant organ or plant. In addition, the present invention provides a recombinant vector comprising an antisense sequence of a nucleic acid sequence encoding pyruvate decarboxylase (PDC), the E1α subunit of pPDH, the E1β subunit of pPDH, the E2 subunit of pPDH, mitochondrial pyruvate dehydrogenase (mtPDH) or aldehyde dehydrogenase (ALDH) or a ribozyme that can cleave an RNA molecule encoding PDC, E1α pPDH, E1β pPDH, E2 pPDH, mtPDH or ALDH.
An isolated or purified nucleic acid molecule encoding a plant plastidic acetyl CoA synthetase (ACS) or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated or purified nucleic acid molecule encoding a modified plant plastidic ACS, which comprises one or more insertions, deletions and/or substitutions, wherein the modified plant plastidic ACS encoded by said isolated or purified nucleic acid molecule does not differ functionally from the corresponding unmodified plant plastidic ACS, or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated or purified nucleic acid molecule encoding the E3 subunit of a plant pPDH or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated or purified nucleic acid molecule encoding a modified E3 subunit of a plant pPDH, which comprises one or more insertions, deletions and/or substitutions, wherein the modified E3 subunit of a plant pPDH encoded by said isolated or purified nucleic acid molecule does not differ functionally from the corresponding unmodified E3 subunit of a plant pPDH, or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated or purified nucleic acid molecule encoding the A subunit of a plant ATP-citrate lyase (ACL) or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated or purified nucleic acid molecule encoding a modified A subunit of a plant ACL, which comprises one or more insertions, deletions and/or substitutions, wherein the modified A subunit of a plant ACL encoded by said isolated or purified nucleic acid molecule does not differ functionally from the corresponding unmodified A subunit of a plant ACL, or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated or purified nucleic acid molecule encoding the B subunit of a plant ACL or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated or purified nucleic acid molecule encoding a modified B subunit of a plant ACL, which comprises one or more insertions, deletions and/or substitutions, wherein the modified B subunit of a plant ACL encoded by said isolated or purified nucleic acid molecule does not differ functionally from the corresponding unmodified B subunit of a plant ACL, or a continuous fragment thereof comprising at least about 20 nucleotides.
An isolated and purified nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:
An isolated and purified nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:
An isolated and purified nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:
A vector comprising or encoding an antisense sequence of at least about 20 nucleotides that hybridizes to or a ribozyme that cleaves an RNA molecule encoding a plant plastidic ACS, the E1α subunit of a plant pPDH, the E1 β subunit of a plant pPDH, the E2 subunit of a plant pPDH, the E3 subunit of a plant pPDH, the A subunit of a plant ACL, the B subunit of a plant ACL, a plant plastidic pyruvate decarboxylase, a plant mitochondrial pyruvate dehydrogenase, or a plant aldehyde dehydrogenase.
A polyclonal antibody that binds to a plant ACS but does not bind to a nonplant ACS.
A polyclonal antibody that binds to the E1α subunit of a plant pPDH but does not bind to a nonplant pyruvate dehydrogenase.
A polyclonal antibody that binds to the E1β subunit of a plant pPDH but does not bind to a nonplant pyruvate dehydrogenase.
A polyclonal antibody that binds to the A subunit of a plant ACL but does not bind to the A subunit of a nonplant ACL.
A polyclonal antibody that binds to the B subunit of a plant ACL but does not bind to the B subunit of a nonplant ACL.
An antisense molecule of at least about 20 nucleotides that can hybridize to an RNA molecule encoding a plant plastidic ACS, the E E1α subunit of a plant pPDH, the E1β subunit of a plant pPDH, the E2 subunit of a plant pPDH, the E3 subunit of a plant pPDH, the A subunit of a plant ACL, the B subunit of a plant ACL, a plant pyruvate decarboxylase, a plant mitochondrial pyruvate dehydrogenase or a plant aldehyde dehydrogenase.
A ribozyme that can cleave an RNA molecule encoding a plant plastidic ACS, the E1α subunit of a plant pPDH, the E1 β subunit of a plant pPDH, the E2 subunit of a plant pPDH, the E3 subunit of a plant pPDH, the A subunit of a plant ACL, the B subunit of a plant ACL, a plant pyruvate decarboxylase, a plant mitochondrial pyruvate dehydrogenase or a plant aldehyde dehydrogenase.
A method of altering the level of an enzyme in a plant cell, a plant tissue, a plant organ, or a plant, which method comprises contacting said plant cell, plant tissue, plant organ or plant with a vector comprising a nucleic acid molecule selected from the group consisting of (i) a gene encoding an enzyme or, if the enzyme is comprised of subunits, a subunit of an enzyme selected from the group consisting of plastidic ACS, pPDH, ACL, pyruvate decarboxylase, acetyl CoA hydrolase, mitochondrial pyruvate dehydrogenase, and aldehyde dehydrogenase, (ii) a nucleic acid molecule comprising or encoding an antisense sequence of at least about 20 nucleotides to an RNA molecule transcribed from a gene of (i), and (iii) a nucleic acid molecule comprising or encoding a ribozyme to an RNA molecule transcribed from a gene of (i), wherein said vector comprising a nucleic acid molecule of (i) increases or decreases the level of said enzyme in said plant cell, plant tissue, plant organ or plant, and wherein said vector comprising a nucleic acid molecule of (ii) or (iii) decreases the level of said enzyme in said plant cell, plant tissue, plant organ or plant.
Dependencies are inferred from the claim text.