Use of a maize untranslated region for transgene expression in plants
View Patent ↗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.
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.