U6 polymerase III promoter and methods of use
AssigneeE. I. Du Pont de Nemours and Company
InventorsZhongsen Li, Zhan-Bin Liu
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Abstract
The disclosure relates to gene expression regulatory sequences from maize, specifically to the promoter of a maize U6 polymerase III gene and fragments thereof and their use in promoting the expression of one or more heterologous nucleic acid fragments in plants. The invention further discloses compositions, polynucleotide constructs, transformed host cells, mutated plants, transgenic plants and seeds containing the recombinant construct with the promoter, and methods for preparing and using the same.
Claims18
A recombinant DNA construct comprising a promoter nucleotide sequence comprising a region of homology of at least 98% sequence identity with at least 100 contiguous nucleotides of SEQ ID NO:
- 9, operably linked to at least one heterologous sequence, wherein said nucleotide sequence controls the expression of the heterologous sequence, wherein the at least 100 contiguous nucleotides comprise a TATA box, and wherein the nucleotide sequence comprises said TATA box.
A method of expressing a coding sequence or a functional RNA in a plant comprising:
- (a) introducing a recombinant DNA construct into the plant wherein the recombinant DNA construct comprises a promoter nucleotide sequence comprising a region of homology of at least 98% sequence identity with at least 100 contiguous nucleotides of SEQ ID NO:
- 9, operably linked to at least one heterologous sequence, wherein said nucleotide sequence controls the expression of the heterologous sequence, wherein the at least 100 contiguous nucleotides comprise a TATA box, wherein the promoter nucleotide sequence comprises said TATA box, and wherein the at least one heterologous sequence comprises a coding sequence or encodes a functional RNA;
- (b) growing the plant of step (a);
- and
- (c) selecting a plant displaying expression of the coding sequence or the functional RNA of the recombinant DNA construct.
A method of transgenically altering a plant trait, comprising:
- (a) introducing a recombinant DNA construct into the plant, wherein the recombinant DNA construct comprises a promoter nucleotide sequence comprising a region of homology of at least 98% sequence identity with at least 100 contiguous nucleotides of SEQ ID NO:
- 9, operably linked to at least one heterologous sequence, wherein said promoter nucleotide sequence controls the expression of the heterologous sequence, wherein the at least 100 contiguous nucleotides comprise a TATA box, and wherein the promoter nucleotide sequence comprises said TATA box;
- (b) growing a fertile, mature plant resulting from step (a);
- and
- (c) selecting a plant expressing the at least one heterologous sequence in at least one plant tissue having the altered trait.
A method for increasing expression of at least one heterologous sequence in a plant comprising:
- (a) transforming a plant cell with a recombinant DNA construct, wherein the recombinant DNA construct comprises a promoter nucleotide sequence comprising a region of homology of at least 98% sequence identity with at least 100 contiguous nucleotides of SEQ ID NO:
- 9, operably linked to at least one heterologous sequence, wherein said promoter nucleotide sequence controls the expression of the heterologous sequence, wherein the at least 100 contiguous nucleotides comprise a TATA box, wherein the promoter nucleotide sequence comprises said TATA box;
- (b) growing fertile mature plants from transformed plant cell of step (a);
- and
- (c) selecting plants containing the transformed plant cell wherein the expression of the heterologous sequence is increased.
A plant transformed with a recombinant DNA construct comprising a promoter and a heterologous nucleic acid fragment operably linked to said promoter, wherein said promoter controls expression of said heterologous nucleic acid fragment in a plant cell, and further wherein said promoter comprises a region of homology of at least 98% sequence identity with at least 100 contiguous nucleotides SEQ ID NO:
- 9;
- wherein the at least 100 contiguous nucleotides comprise a TATA box, and wherein the promoter comprises said TATA box.
A recombinant DNA construct comprising a promoter comprising a nucleotide sequence comprising a region of homology of at least 98% sequence identity with at least 100 contiguous nucleotides with SEQ ID NO:
- 9, wherein the at least 100 contiguous nucleotides comprise a TATA box, wherein the nucleotide sequence comprises said TATA box, wherein said promoter is operably linked to a heterologous nucleic acid fragment, and wherein said heterologous nucleic acid fragment expresses a guide RNA.
Dependencies are inferred from the claim text.