Regulation of quinolate phosphoribosyl transferase expression
AssigneeNorth Carolina State University
InventorsMark A. Conkling, Wen Song, Nandini Mendu
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Abstract
DNA encoding a plant quinolate phosphoribosyl transferase (QPRTase) enzyme, and constructs comprising such DNA are provided. Methods of altering quinolate phosphoribosyl transferase expression are provided.
Claims93
An isolated DNA molecule comprising a sequence selected from the group consisting of:
- (a) SEQ ID NO:1;
- (b) DNA sequences which encode an enzyme having SEQ ID NO:2;
- (c) DNA sequences which hybridize to isolated DNA of (a) or (b) above and which encode a quinolate phosphoribosyl transferase enzyme;
- and
- (d) DNA sequences which differ from the DNA of (a), (b) or (c) above due to the degeneracy of the genetic code.
A DNA construct comprising, in the 5′ to 3′ direction, a promoter operable in a plant cell and DNA encoding a plant quinolate phosphoribosyl transferase, said DNA operably associated with said promoter.
A DNA construct comprising, in the 5′ to 3′ direction, a promoter operable in a plant cell and DNA encoding a plant quinolate phosphoribosyl transferase, said DNA in antisense orientation and operably associated with said promoter.
A peptide having SEQ ID NO:2.
A peptide encoded by a DNA sequence selected from the group consisting of:
- (a) SEQ ID NO:1;
- (b) DNA sequences which hybridize to isolated DNA of (a) above and which encode a quinolate phosphoribosyl transferase enzyme;
- and
- (c) DNA sequences which differ from the DNA of (a) or (b) above due to the degeneracy of the genetic code.
A method of making a transgenic plant cell having reduced quinolate phosphoribosyl transferase (QPRTase) expression, said method comprising:
- providing a plant cell of a type known to express quinolate phosphoribosyl transferase;
- providing an exogenous DNA construct, which construct comprises, in the 5′ to 3′ direction, a promoter operable in a plant cell and DNA comprising a portion of a sequence encoding quinolate phosphoribosyl transferase mRNA, said DNA operably associated with said promoter;
- and
- transforming said plant cell with said DNA construct to produce transformed cells, said plant cell having reduced expression of QPRTase compared to an untransformed cell.
A method of producing transgenic tobacco seeds, comprising collecting seed from a transgenic tobacco plant produced by the method of claim 32.
A transgenic plant of the species Nicotiana having reduced quinolate phosphoribosyl transferase (QPRTase) expression relative to a non-transformed control plant, said transgenic plant comprising transgenic plant cells containing:
- an exogenous DNA construct comprising, in the 5′ to 3′ direction, a promoter operable in said plant cell and DNA comprising a segment of a DNA sequence that encodes a plant quinolate phosphoribosyl transferase mRNA, said DNA operably associated with said promoter;
- said plant exhibiting reduced QPRTase expression compared to a non-transformed control plant.
A method for reducing expression of a quinolate phosphoribosyl transferase gene in a plant cell, said method comprising:
- growing a plant cell transformed to contain exogenous DNA, wherein a transcribed strand of said exogenous DNA is complementary to quinolate phosphoribosyl transferase mRNA endogenous to said cell, whereby transcription of said complementary strand reduces expression of said quinolate phosphoribosyl gene.
A method of producing a tobacco plant having decreased levels of nicotine in leaves of said tobacco plant, said method comprising:
- growing a tobacco plant, or progeny plants thereof, wherein said plant comprises cells containing a DNA construct comprising a transcriptional initiation region functional in said plant and an exogenous DNA sequence operably joined to said transcriptional initiation region, wherein a transcribed strand of said DNA sequence is complementary to endogenous quinolate phosphoribosyl transferase messenger RNA in said cells.
A method of making a transgenic plant cell having increased quinolate phosphoribosyl transferase (QPRTase) expression, said method comprising:
- providing a plant cell of a type known to express quinolate phosphoribosyl transferase;
- providing an exogenous DNA construct, which construct comprises, in the 5′ to 3′ direction, a promoter operable in a plant cell and a DNA sequence encoding quinolate phosphoribosyl transferase, said DNA sequence operably associated with said promoter;
- and
- transforming said plant cell with said DNA construct to produce transformed cells, said plant cell having increased expression of QPRTase compared to an untransformed cell.
A transgenic plant of the species Nicotiana having increased quinolate phosphoribosyl transferase (QPRTase) expression relative to a non-transformed control plant, said transgenic plant comprising transgenic plant cells containing:
- an exogenous DNA construct comprising, in the 5′ to 3′ direction, a promoter operable in said plant cell and a DNA sequence encoding a plant quinolate phosphoribosyl transferase, said DNA operably associated with said promoter;
- said plant exhibiting increased QPRTase expression compared to a non-transformed control plant.
A method for increasing expression of a quinolate phosphoribosyl transferase gene in a plant cell, said method comprising:
- growing a plant cell transformed to contain exogenous DNA, wherein said exogenous DNA encodes quinolate phosphoribosyl transferase.
A method of producing a tobacco plant having increased levels of nicotine in leaves of said tobacco plant, said method comprising:
- growing a tobacco plant, or progeny plants thereof, wherein said plant comprises cells containing a DNA construct comprising a transcriptional initiation region functional in said plant and an exogenous DNA sequence operably joined to said transcriptional initiation region, wherein said DNA sequence encodes quinolate phosphoribosyl transferase functional in said cells.
Dependencies are inferred from the claim text.