Seed-Preferred Regulatory Elements

a technology of seed-based elements and regulatory elements, applied in the field of plant molecular biology, can solve problems such as the influence of one or more genes on the expression of genes

Inactive Publication Date: 2009-02-19
PIONEER HI BRED INT INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0007]The invention is directed to a promoter from a Sorghum bicolor oleosin-encoding gene, useful as a regulatory region and providing for expression of a nucleotide sequence of interest. In an embodiment the expression is driven in a seed-embryo/aleurone preferred manner. The invention is further directed to functional fragments w

Problems solved by technology

Diverse regulatory sequences are also needed as undesirable biochemical interactions can result from using the same regulatory sequence to control more

Method used

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  • Seed-Preferred Regulatory Elements
  • Seed-Preferred Regulatory Elements

Examples

Experimental program
Comparison scheme
Effect test

example 1

Isolation of Regulatory Sequences

[0068]Regulatory regions from sorghum SB-OLE (Sorghum bicolor oleosin protein) were isolated from sorghum plants and cloned. Sorghum SB-OLE was selected as a source of seed preferred regulatory elements based on the spatial and temporal expression of its products.

[0069]The promoter is a 948 base pair nucleotide sequence, shown in FIG. 1 with the TATA box underlined. A BLAST of GenBank showed the highest identity was 59% to GenBank accession number U13701 (GI:687244) by Lee and Huang, “Zea mays oil body protein 16 kDa oleosin (ole16) gene, complete cds” 1995; and Lee and Huang (1994), supra. A region of 433 base pairs in the 3′ region of the sequence indicates 78% identity. A comparison is shown in FIG. 2 of the Zea mays oleosin promoter (SEQ ID NO: 3, the Sorghum oleosin promoter (SEQ ID NO: 1) and the Oryza sativa (SEQ ID NO: 2) oleosin promoter.

[0070]Motifs were identified that were common to all three oleosin promoters, thus indicating the importa...

example 2

Expression Data Using Promoter Sequences

[0075]A construct, named PHP28986, was prepared which included the sorghum oleosin promoter (SEQ ID NO:1) linked with the YFP selectable marker, supra and the nos or nopaline synthase transcription terminator (Depicker et al. (1982) J. Mol. Appl. Genet. 1(6):561-573; Shaw et al. (1984) Nucleic Acids Research Vol. 12, No. 20, pp. 7831-7846). All vectors were constructed using standard molecular biology techniques (Sambrook et al., supra). Successful subcloning was confirmed by restriction analysis.

example 3

Transformation and Regeneration of Maize Callus Via Agrobacterium

[0076]Constructs used were as those set forth supra using a binary plasmid with the left and right borders (see Hiei et al., U.S. Pat. No. 7,060,876) and the selectable marker for maize-optimized PAT (phosphinothricin acetyl transferase). Jayne et al., U.S. Pat. No. 6,096,947.

Preparation of Agrobacterium Suspension:

[0077]Agrobacterium was streaked out from a −80° frozen aliquot onto a plate containing PHI-L medium and was cultured at 28° C. in the dark for 3 days. PHI-L media comprises 25 ml / l Stock Solution A, 25 ml / l Stock Solution B, 450.9 ml / l Stock Solution C and spectinomycin (Sigma Chemicals) was added to a concentration of 50 mg / l in sterile ddH20 (stock solution A: K2HPO4 60.0 g / l, NaH2PO4 20.0 g / l, adjust pH to 7.0 w / KOH and autoclaved; stock solution B: NH4Cl 20.0 g / l, MgSO4.7H2O 6.0 g / l, KCl 3.0 g / l, CaCl2 0.20 g / l, FeSO4.7H2O 50.0 mg / l, autoclaved; stock solution C: glucose 5.56 g / l, agar 16.67 g / l (#A-70...

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Abstract

The present invention provides compositions and methods for regulating expression of nucleotide sequences in a plant. Compositions are novel nucleotide sequences for a tissue preferred promoter isolated from the sorghum 16 kDa oleosin coding region. The sequences drive expression preferentially to seed tissue, and most preferably to embryo and/or aleurone tissue of a plant. Functional fragments of same are also provided. A method for expressing a nucleotide sequence in a plant using the regulatory sequences disclosed herein is provided. The method comprises transforming a plant cell to comprise a nucleotide sequence operably linked to one or more of the regulatory sequences of the present invention and regenerating a stably transformed plant from the transformed plant cell.

Description

CROSS-REFERENCE PARAGRAPH[0001]This application claims the benefit of U.S. Provisional Application No. 60 / 955,409, filed Aug. 13, 2007, which is herein incorporated by reference in its entirety.FIELD OF THE INVENTION[0002]The present invention relates to the field of plant molecular biology, more particularly to regulation of gene expression in plants.BACKGROUND OF THE INVENTION[0003]Expression of DNA sequences in a plant host is dependent upon the presence of regulatory elements that are functional within the plant host. Choice of the regulatory element will determine when and where within the organism the DNA sequence is expressed. Where continuous expression is desired throughout the cells of a plant, and / or throughout development, constitutive promoters are utilized. In contrast, where gene expression in response to a stimulus is desired, inducible promoters are the regulatory element of choice. Where expression in specific tissues or organs are desired, tissue-preferred promote...

Claims

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Application Information

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IPC IPC(8): C12N15/11A01H5/00
CPCC12N15/8234C07K14/415
Inventor ABBITT, SHANE E.
Owner PIONEER HI BRED INT INC
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