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AssigneeHer Majesty the Queen in Right of Canada as Rep.by the Minister of Agriculture and Agri-Food Canada
InventorsBrian Miki, Therese Ouellet, Jiro Hattori, Elizabeth Foster, Helene Labbe, Teresa Martin-Heller, Lining Tian, Daniel Charles William Brown, Peijun Zhang, Keqiang Wu
Estimating the term…
T-DNA tagging with a promoterless β-glucuronidase (GUS) gene generated transgenic Nicotiana tabacum plants that expressed GUS activity either only in developing seed coats, or constitutively. Cloning and deletion analysis of the GUS fusion revealed that the promoter responsible for seed coat specificity was located in the plant DNA proximal to the GUS gene. Analysis of the region demonstrated that the seed coat-specificity of GUS expression in this transgenic plant resulted from T-DNA insertion next to a cryptic promoter. This promoter is useful in controlling the expression of genes to the developing seed coat in plant seeds. Similarly, cloning and characterization of the cryptic constitutive promoter revealed the occurrence of several cryptic regulatory regions. These regions include promoter, negative regulatory elements, transcriptional enhancers, core promoter regions, and translational enhancers and other regulatory elements.
1 An isolated nucleic acid comprising a cryptic regulatory element obtained from a plant.
The cryptic regulatory element of claim 1, said cryptic regulatory element comprising a promoter.
The cryptic regulatory element of claim 1, said cryptic regulatory element comprising a core promoter.
The cryptic regulatory element of claim 1, said cryptic regulatory element comprising an enhancer.
The cryptic regulatory element of claim 1, said cryptic regulatory element comprising a negative regulatory element.
The cryptic regulatory element of claim 1, said cryptic regulatory element comprising a post-transcriptional regulatory element.
The cryptic regulatory element of claim 1, wherein said cryptic regulatory element is selected from the group consisting of a tissue-specific regulatory element and a constitutive regulatory element.
The cryptic regulatory element of claim 1, comprising a DNA fragment of about 2.5 kb and characterized by the restriction map of FIG. 2(B).
A vector comprising the cryptic regulatory element as defined in claim 1, operatively associated with a gene that encodes a protein, wherein the gene is under the control of said cryptic regulatory element.
A transgenic plant containing a cryptic regulatory element as claimed in claim 1, operatively linked to a gene encoding a protein.
21 The cryptic regulatory element of claim 12, comprising an XbaI and SnaBI fragment of about 1.5 kb.
A seed obtained from a transgenic plant containing a cryptic regulatory element as claimed in claim 1, operatively linked to a gene encoding a protein.
37 The cryptic regulatory element of claim 29, comprising nucleotides 1660-2086 of SEQ ID NO:2
39 The cryptic regulatory element of claim 29, comprising nucleotides 1875-2086 of SEQ ID NO:2
A method of conferring expression of a gene in a host organism, comprising operatively linking an exogenous DNA of interest, for which expression is desired with the cryptic regulatory element of claim 1, or a fragment thereof, to produce a chimeric gene construct, and introducing the chimeric gene construct into the host organism capable of expressing the chimeric gene construct.
A method of modulating expression of a gene in a host organism, comprising operatively linking an exogenous DNA of interest, for which expression is desired with a promoter of interest and the cryptic regulatory element of claim 1, or a fragment thereof, to produce a chimeric gene construct, and introducing the chimeric gene construct into the host organism capable of expressing the chimeric gene construct.
A method of modulating expression of a gene in a host organism, comprising operatively linking an exogenous DNA of interest, for which expression is desired with a promoter of interest and at least one fragment of the cryptic regulatory element of claim 1 to produce a chimeric gene construct, and introducing the chimeric gene construct into the host organism capable of expressing the chimeric gene construct.
65 The method of claim 40, wherein the exogenous DNA of interest encodes a protein selected from the group consisting of a pharmaceutically active protein, a growth factor, a growth regulator, an antibody, an antigen, an interleukin, insulin, G-CSF, GM-CSF, hPG-CSF, M-CSF, an interferon, a blood clotting factor, an industrial enzyme, a protein supplement, a nutraceutical, a protease, an oxidases, a phytase, a chitinase, an invertase, a lipase, a cellulase, a xylanase, and an enzyme involved in oil metabolic and biosynthetic pathways.
A transgenic host organism containing a cryptic regulatory element as claimed in claim 1, operatively linked to a gene encoding a protein.
Dependencies are inferred from the claim text.