Root-preferred promoter from a brachypodium distachyon metallothionein-like gene

Patent metadata

Field	Value
Publication number	US-10443067-B2
Application number	US-201715835595-A
Country	US
Kind code	B2
Filing date	Dec 8, 2017
Priority date	Apr 15, 2015
Publication date	Oct 15, 2019
Grant date	Oct 15, 2019

Abstract

This disclosure concerns compositions and methods for promoting transcription of a nucleotide sequence in a plant or plant cell, employing a promoter from a Brachypodium distachyon metallothionein-like gene (mtl). Some embodiments relate to a promoter from a Brachypodium distachyon metallothionein-like gene (mtl) that functions in plants to promote transcription of operably linked nucleotide sequences.

First claim

What is claimed is:

1. A gene expression cassette comprising a promoter operably linked to a heterologous nucleic acid, wherein the promoter comprises a polynucleotide comprising a sequence identity of at least 97% to SEQ ID NO:1, and said promoter has below ground tissue preferred expression.

2. The gene expression cassette of claim 1 , wherein the polynucleotide further comprises an intron.

3. The gene expression cassette of claim 1 , wherein the polynucleotide further comprises a 5′ UTR.

4. The gene expression cassette of claim 1 , wherein the polynucleotide is operably linked to a 3′ UTR.

5. The gene expression cassette of claim 4 , wherein the 3′ UTR comprises SEQ ID NO:3.

6. The gene expression cassette of claim 1 , wherein the operably linked heterologous nucleic acid encodes a polypeptide or a small RNA gene.

7. The gene expression cassette of claim 1 , wherein the heterologous nucleic acid is selected from the group consisting of a heterologous nucleic acid conferring insecticidal resistance, a heterologous nucleic acid conferring herbicide tolerance, a heterologous nucleic acid conferring nitrogen use efficiency, a heterologous nucleic acid conferring water use efficiency, a heterologous nucleic acid conferring nutritional quality, a heterologous nucleic acid encoding a DNA binding protein, and a heterologous nucleic acid encoding a selectable marker.

8. A recombinant vector comprising the gene expression cassette of claim 1 , wherein the vector is selected from the group consisting of a plasmid, a cosmid, a bacterial artificial chromosome, a virus, and a bacteriophage.

9. A transgenic cell comprising the gene expression cassette of claim 1 .

10. The transgenic cell of claim 9 , wherein the transgenic cell is a transgenic plant cell.

11. A transgenic plant comprising the transgenic plant cell of claim 10 .

12. The transgenic plant of claim 11 , wherein the transgenic plant is a monocotyledonous plant or a dicotyledonous plant.

13. The transgenic plant of claim 12 , wherein the monocotyledonous plant is selected from the group consisting of a maize plant, a rice plant, and a wheat plant.

14. A transgenic seed from the transgenic plant of claim 12 , wherein the seed comprises the gene expression cassette.

15. The gene expression cassette of claim 1 , wherein the promoter comprises the polynucleotide sequence of nucleotides 1 to 2,000 of SEQ ID NO:1.

16. A method for expressing a coding sequence in a transgenic plant, the method comprising:
a) transforming a plant cell with a gene expression cassette comprising a polynucleotide sequence comprising a sequence identity of at least 97% to SEQ ID NO:1 operably linked to a heterologous coding sequence, which is operably linked to a 3′ untranslated region;
b) isolating the transformed plant cell comprising the gene expression cassette;
c) regenerating a transgenic plant from the transformed plant cell; and,
d) obtaining the transgenic plant, wherein the transgenic plant expresses the coding sequence with below ground tissue preferred expression.

17. A method for manufacturing a synthetic polynucleotide sequence comprising a sequence identity of at least 97% to SEQ ID NO:1, the method comprising:
a) isolating a nucleic acid comprising a polynucleotide sequence comprising SEQ ID NO:1;
b) producing a plurality of oligonucleotide primer sequences, wherein the oligonucleotide primer sequences bind to the nucleic acid under stringent hybridization conditions;
c) ligating the plurality of oligonucleotide primer sequences to synthesize a synthetic polynucleotide sequence; and,
d) sequencing the resulting synthetic polynucleotide to confirm that it comprises at least 97% identity to SEQ ID NO:1.

Assignees

• Dow Agrosciences Llc

Inventors

• Kumar Sandeep
• Hemingway Daren
• Ausmus Carla
• Worden Andrew F
• Asberry Andrew

Classifications

• C12N15/8227 Primary

  Root-specific · CPC title

• Y02A40/146

  Genetically Modified [GMO] plants, e.g. transgenic plants · CPC title

• C12N15/8216

  Methods for controlling, regulating or enhancing expression of transgenes in plant cells · CPC title

• C07K14/825

  Metallothioneins · CPC title