Control of gene expression in plants by receptor mediated transactivation in the presence of a chemical ligand

• US 6,939,711 B2
• Filed: 03/21/2001
• Issued: 09/06/2005
• Est. Priority Date: 03/03/1995
• Status: Expired due to Fees

• Current Assignee: SYNGENTA INVESTMENT CORPORATION
• Original Assignee: SYNGENTA INVESTMENT CORPORATION
• Inventors: Privalle, Laura S., Goff, Stephen A., Crossland, Lyle D.
• Primary Examiner(s): WOITACH, JOSEPH T
• Application Number:
  US09/813,589
• Publication Number:
  US 20010022004A1
• Time in Patent Office: 1,630 Days
• Field of Search: 435/320.1, 435/419, 534/23.1, 534/23.4, 534/24.1, 534/24.5, 800/288
• US Class Current: 435/419
• CPC Class Codes: C07K 14/70567 Nuclear receptors, e.g. retinoic acid receptor [RAR], RXR, nuclear orphan receptors
• CPC Class Codes: C07K 14/721 Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/8217 Gene switch
• CPC Class Codes: C12N 15/8237 Externally regulated expression systems
• CPC Class Codes: C12N 15/8289 Male sterility
• CPC Class Codes: C12N 15/829 Female sterility

Abstract

The present invention is drawn to a method of controlling gene expression in plants. Specifically, the method comprises obtaining a transgenic plant comprising at least two receptor expression cassettes and at least one target expression cassette. The first receptor expression cassette comprises a nucleotide sequence for a 5′ regulatory region operably linked to a nucleotide sequence which encodes a first receptor polypeptide, and a 3′ termination region. The second receptor expression cassette comprises a nucleotide sequence for a 5′ regulatory region operably linked to a nucleotide sequence which encodes a second receptor polypeptide, and a 3′ termination region. The target expression cassette comprises a nucleotide sequence for a 5′ regulatory region operably linked to a nucleotide sequence which encodes a target polypeptide, and a 3′ termination region, wherein the 5′ regulatory region of the target expression cassette is activated by the first and second receptor polypeptides in the presence of one or more chemical ligands which are complementary to the ligand binding domain of the receptor polypeptides, whereby expression of the target polypeptide is accomplished. The method is useful for controlling various traits of agronomic importance, such as plant fertility.

Claims

1. A receptor expression cassette for use in a method of controlling gene expression in a plant, comprising:

(a) a 5′

regulatory region capable of promoting expression in a plant cell;

(b) an operably linked coding sequence encoding an Ecdysone receptor polypeptide comprising a ligand binding domain and a DNA binding domain, and a heterologous transactivation domain, wherein said heterologous transactivation domain is the transactivation domain from the C1 regulatory gene of maize; and

(c) a 3′

terminating sequence.

2. A plant transformation vector comprising the receptor expression cassette of claim 1.

3. A plant cell transformed with the plant transformation vector of claim 2.