Transcription regulating nucleotide sequences and methods of use
AssigneeBASF Agricultural Solutions Seed US LLC
InventorsErin Marie Davis, Sebastian Hermann Martschat, Jonathan T. Vogel, Hunter James Cameron, Zhixin Shi
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Abstract
Described herein are transcription regulating nucleotide sequences and the use of such transcription regulating nucleotide sequences to express a polynucleotide of interest in plants.
Claims66
A transcription regulating nucleotide sequence that confers constitutive gene expression in a plant, wherein said transcription regulating nucleotide sequence has been modified by deleting at least one occurrence of the Motif from its DNA sequence.
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An expression cassette for regulating constitutive expression of a polynucleotide of interest, said expression cassette comprising a transcription regulating nucleotide sequence having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, 100% sequence identity to any one of the nucleic acid sequences set forth in SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, SEQ ID NO:
- 5, SEQ ID NO:
- 6, or a functional fragment thereof.
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A method of identifying a promoter, the method comprising the steps of:
- (a) analyze RNA sequence expression datasets across tissue types from a plant species;
- (b) identify at least one k-mer associated with the desired gene expression;
- (c) identify a CDS having said at least one k-mer upstream of said CDS;
- (d) select at least 100 bp, 200 bp, 300 bp, 400 bp, 500 bp, 600 bp, 700 bp, 800 bp, 900 bp or greater than or equal to 1000 bp, 1500 bp, or 2000 bp nucleotide sequence upstream from said CDS:
- thereby identifying a promoter sequence.
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A promoter that has been modified from its native form by (a) deleting at least one occurrence of the sequence GATCTG;
- or (b) modifying the Motif sequence wherein such deletion or modification results in promoter having a decreased expression variance as compared to the same promoter that does not contain said deletion or modification.
A method of making a promoter with decreased expression variance, the method comprising the steps of:
- (a) analyze RNA sequence expression datasets across tissue types from a plant species;
- (b) identify at least one k-mer associated with the desired gene expression;
- (c) identify a CDS having said at least one k-mer upstream of said CDS;
- (d) select at least 100 bp, 200 bp, 300 bp, 400 bp, 500 bp, 600 bp, 700 bp, 800 bp, 900 bp or greater than or equal to 1000 bp nucleotide sequence upstream from said CDS;
- (e) remove or modify the sequence of at least one instance of said at least one k-mer identified in step (b);
- thereby making a promoter with decrease expression variance.
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A expression cassette comprising a soybean constitutive promoter having at least one instance of the k-mer GATCTG.
A plant expression vector comprising a promoter operably linked to a gene of interest having at least one instance of the k-mer GATCTG.
A soybean promoter whose native form has been modified by editing or deleting at least one, at least 2, or at least 3, or more instances of the k-mer GATCTG.
Dependencies are inferred from the claim text.