Use of a maize untranslated region for transgene expression in plants

Patent metadata

Field	Value
Publication number	US-9688996-B2
Application number	US-201314416811-A
Country	US
Kind code	B2
Filing date	Aug 8, 2013
Priority date	Aug 17, 2012
Publication date	Jun 27, 2017
Grant date	Jun 27, 2017

Abstract

Provided are methods, vectors and gene constructs for enhancing expression of a recombinant nucleic acid sequence in transgenic plants and plant tissues. According to the present invention, nucleic acid sequences are obtained and/or derived from the 3′ untranslated regions of genes encoding ubiquitin proteins and engineered to flank respective portions of a selected coding region of a vector. The vector construct may be introduced into plants and/or plant tissues through conventional or gene targeting procedures, resulting in enhanced expression of the selected coding region. In some embodiments, the selected coding region is a chimeric gene or gene fragment expressing one or more proteins known to impart a level of insecticidal activity to a transgenic plant and/or plant tissue.

First claim

We claim:

1. A nucleic acid construct comprising at least one structural gene of interest functionally linked both to a heterologous promoter and one or more control sequences selected from the group consisting of SEQ ID NOS: 1-3, their full complements, and combinations thereof.

2. The nucleic acid construct of claim 1 , wherein the at least one structural gene of interest comprises a gene that confers a non-native phenotype in a plant.

3. The nucleic acid construct of claim 1 , wherein the at least one structural gene of interest comprises a gene that confers insect resistance or herbicide resistance/tolerance in a plant.

4. The nucleic acid construct of claim 1 , wherein the one or more control sequences are amplified using oligonucleotides selected from the group consisting of SEQ ID NOS: 4-15.

5. The nucleic acid construct of claim 1 , wherein the nucleic acid construct comprises a binary vector for Agrobacterium -mediated transformation.

6. The nucleic acid construct of claim 1 , wherein the nucleic acid construct is stably transformed into a transgenic plant.

7. The nucleic acid construct of claim 6 , wherein the transgenic plant is a monocotyledon plant.

8. The nucleic acid construct of claim 6 , wherein the transgenic plant is a dicotyledon plant.

9. The nucleic acid construct of claim 1 , wherein the nucleic acid construct comprises a selectable marker.

10. The nucleic acid construct of claim 9 , wherein the selectable marker comprises an aryloxyalkanoate dioxygenase.

11. The nucleic acid construct of claim 10 , wherein the aryloxyalkanoate dioxygenase is AAD-1 or AAD-12.

12. A vector comprising the nucleic acid construct of claim 1 .

13. A plant or plant cell transformed with the nucleic acid construct of claim 1 .

14. The plant or plant cell of claim 13 further comprising an additional structural gene of interest stacked with the at least one gene of interest.

15. A method for recombinantly producing a peptide or protein comprising functionally linking to a gene encoding the peptide or protein both a heterologous promoter and one or more control sequences selected from the group consisting of SEQ ID NOS: 1-3, their full complements, and combinations thereof.

16. The method of claim 15 , wherein the one or more control sequences are amplified using oligonucleotides selected from the group consisting of SEQ ID NOS: 4-15.

17. A method for expression of a gene in a plant or plant cell comprising functionally linking to the gene both a heterologous promoter and one or more control sequences selected from the group consisting of SEQ ID NOS: 1-3, their full complements, and combinations thereof.

18. The method of claim 17 , wherein the one or more control sequences are amplified using oligonucleotides selected from the group consisting of SEQ ID NOS: 4-15.

Assignees

• Dow Agrosciences Llc
• Kumar Sandeep
• Gupta Manju
• Alabed Diaa

Inventors

• Kumar Sandeep
• Gupta Manju
• Alabed Diaa

Classifications

• C12N15/8216 Primary

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

• C12N15/8218 Primary

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• C12N15/8286

  for insect resistance · CPC title

• C12N15/8274

  for herbicide resistance · CPC title

• C12N15/821

  Non-antibiotic resistance markers, e.g. morphogenetic, metabolic markers · CPC title