Plant regulatory sequences for selective control of gene expression
AssigneeMonsanto Technology LLC
InventorsTimothy W. Conner, Patrice Dubois, Marianne Malven, James D. Masucci
Life of this patent
Estimating the term…
Abstract
Promoters from male reproductive tissues are isolated from corn and wheat. These promoters can be used in plants to regulate transcription of target genes including genes for control of fertility, insect or pathogen tolerance, herbicide tolerance or any gene of interest.
Claims22
An isolated nucleic acid comprising a sequence selected from the group consisting of SEQ ID NOs:
- 35-62, or a fragment, region, or cis element of said sequence thereof, said isolated nucleic acid being capable of regulating transcription of an operably linked DNA sequence.
A promoter comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
- 35-62 and fragments thereof.
A cell comprising a recombinant DNA construct comprising an isolated nucleic acid sequence selected from the group consisting of SEQ ID NOs:
- 35-62 or a fragment, region, or cis element of said sequence thereof, and operably linked to said nucleic acid sequence, a transcribable DNA sequence and a 3′ non-translated region.
A transgenic plant comprising a DNA construct comprising an isolated nucleic acid sequence selected from the group consisting of SEQ ID NOs:
- 35-62 or a fragment, region, or cis element of said sequence thereof, and operably linked to said nucleic acid sequence, a transcribable DNA sequence and a 3′ non-translated region.
A method of regulating transcription of a DNA sequence comprising operably linking the DNA sequence to a promoter comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
- 35-62.
A method of making a transgenic plant comprising introducing into a cell of a plant a recombinant DNA construct comprising:
- (i) a promoter comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
- 35-62 or a fragment, region or cis element thereof, and, operably linked to the promoter, (ii) a transcribable DNA sequence and (iii) a 3′ non-translated region.
A method of isolating at least two 5′ regulatory sequences that confer enhanced expression of operably linked genes in male reproductive tissues from a plant comprising:
- (i) evaluating a collection of nucleic acid sequences of ESTs derived from at least one cDNA library prepared from a plant cell type of interest;
- (ii) comparing EST sequences from at least one target plant cDNA library and at least one non-target cDNA libraries of ESTs from a different plant cell type;
- (iii) subtracting common EST sequences found in both target and non-target libraries;
- (iv) designing gene specific primers from the remaining EST sequences after said subtraction;
- and
- (v) isolating the corresponding 5′ flanking and regulatory sequences from a genomic library prepared from the target plant comprising the use of said primers.
A method of isolating at least two 5′ regulatory sequences from cDNA library that confer enhanced expression of operably linked genes in male reproductive tissues from a plant comprising:
- (i) preparing nucleotide sequences from randomly picked a plurality of clones derived from at least one cDNA library prepared from a plant anther tissue;
- (ii) immobilizing said nucleotide sequences onto a piece of glass or a nylon membrane;
- (iii) hybridizing said nucleotide sequences with labeled probes, wherein said labeled probes comprise specific cDNA nucleotide sequences prepared from total mRNA isolated from a known anther plant tissue;
- (iv) identifying nucleotide sequences that are hybridized by said labeled probes;
- (v) designing gene specific primers from said nucleotide sequences of step (iv);
- and
- (vi) isolating the corresponding 5′ flanking and regulatory sequences from said plurality of clones using said primers.
Dependencies are inferred from the claim text.