Methods and compositions for modified ethanol inducible promoter systems

Patent metadata

Field	Value
Publication number	US-9434953-B2
Application number	US-201113978133-A
Country	US
Kind code	B2
Filing date	Dec 15, 2011
Priority date	Jan 18, 2011
Publication date	Sep 6, 2016
Grant date	Sep 6, 2016

Abstract

The present invention provides nucleic acid molecules comprising one or more nucleotide sequences selected from the group consisting of the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:10 that can be operably linked to a promoter, thereby making the promoter inducible by a chemical compound that can induce the expression of the alcohol dehydrogenase system of Aspergillus nidulans . Methods for making an inducible promoter and for making the expression of a nucleotide sequence of interest inducible are also provided. Further provided are plants, plant parts, and plant cells, comprising the compositions of the present invention.

First claim

What is claimed is:

1. An isolated nucleic acid molecule comprising five to nine nucleotide sequences selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 10, wherein the selected nucleotide sequences can be in any combination, in any orientation, and/or in any order, and wherein the five to nine nucleotide sequences are operably linked to a heterologous promoter.

2. The isolated nucleic acid molecule of claim 1 , wherein the heterologous promoter is further operably linked to a nucleotide sequence of interest.

3. A method of making a promoter inducible, comprising:
a) constructing a nucleic acid molecule comprising five to nine nucleotide sequences selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 10, in any combination, in any orientation, and/or in any order; and
b) operably linking the nucleic acid molecule of (a) with the promoter, wherein the promoter is heterologous to the five to nine nucleotide sequences of the nucleic acid molecule.

4. The method of claim 3 , wherein the heterologous promoter is made inducible to a chemical compound selected from the group consisting of a primary alcohol, a primary monoamine, a ketone, a methyl ketone, a hydrolysable ester, an aliphatic aldehyde, and any combination thereof.

5. The method of claim 3 , wherein the heterologous promoter is made inducible to a chemical compound selected from the group consisting of ethanol, acetaldehyde, ethyl methyl ketone, acetone, ethylamine, cyclohexanone, butan-2-ol, 3-oxobutyric acid, propan-2-ol, propan-1-ol, butan-2-ol, threonine, and any combination thereof.

6. The method of claim 3 , wherein the heterologous promoter that is made inducible is a constitutive promoter, a tissue specific promoter, a developmentally regulated promoter, and/or a chemically inducible promoter that is inducible by compounds that do not induce the expression of the alcohol dehydrogenase system of Aspergillus nidulans.

7. The method of claim 3 , wherein the heterologous promoter is a minimal promoter.

8. The method of claim 3 , wherein the five to nine nucleotide sequences are at the 5 ′ end of the nucleic acid molecule.

9. A method of making the expression of a nucleotide sequence of interest inducible comprising operably linking the heterologous promoter of claim 1 to the nucleotide sequence of interest.

10. A promoter made by the method of claim 3 .

11. A plant, plant part or plant cell comprising the isolated nucleic acid molecule of claim 2 .

Assignees

• Sainz Manuel Benito
• Kinkema Mark David
• Syngenta Participations Ag
• Queensland Univ Of Tech

Inventors

• Sainz Manuel Benito
• Kinkema Mark David

Classifications

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N15/8238 Primary

  chemically inducible, e.g. tetracycline · CPC title

• C12N15/8241

  Phenotypically and genetically modified plants via recombinant DNA technology · CPC title