Chimeric gene for the transformation of plants

• US 5,510,471 A
• Filed: 05/31/1994
• Issued: 04/23/1996
• Est. Priority Date: 03/05/1991
• Status: Expired

• Current Assignee: Rhone-Poulenc Agro
• Original Assignee: Rhone-Poulenc Agrochimie
• Inventors: Sailland, Alain, Lebrun, Michel, Leroux, Bernard
• Primary Examiner(s): Fox, David T.
• Assistant Examiner(s): McElwain, Elizabeth F.
• Application Number:
  US08/251,621
• Time in Patent Office: 693 Days
• Field of Search: 800/205, 536/23.4, 536/23.6, 435/172.3, 435/69.7, 435/69.1, 935/11
• US Class Current: 536/23.4
• CPC Class Codes: C07K 2319/08 containing a chloroplast localisation signal
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/8275 Glyphosate

Abstract

Chimeric gene for conferring to plants an increased tolerance to a herbicide having as its target EPSPS comprises, in the direction of transcription, a promoter region, a transit peptide region, a coding sequence for glyphosate tolerance and a polyadenylation signal region, wherein the transit peptide region comprises, in the direction of translation, at least one transit peptide of a plant gene encoding a plastid-localised enzyme, a partial sequence of the N-terminal mature part of a plant gene encoding a plastid-localised enzyme and then a second transit peptide of a plant gene encoding a plastid- localised enzyme. Production of glyphosate-tolerant plants is disclosed.

Claims

1. A nucleic acid construct which codes for a polypeptide sufficient for localization of a gene product in a chloroplast of a plant cell which polypeptide comprises a fusion which in the direction of translation comprises a first chloroplast transit peptide from a ribulose-1,5-bisphosphate carboxylase small subunit, an N-terminal domain of a mature ribulose-1,5-bisphosphate carboxylase small subunit protein and a second chloroplast transit peptide from a ribulose-1,5-bisphosphate carboxylase small subunit.

2. The nucleic acid construct of claim 1 wherein said first chloroplast transit peptide and said second chloroplast transit peptide have identical amino acid sequences.

3. The nucleic acid construct of claim 1 wherein said first chloroplast transit peptide and said second chloroplast transit peptides each have different amino acid sequences.

4. The nucleic acid construct of claim 1 wherein said first or second transit peptide is a transit peptide of a maize ribulose-1,5-bisphosphate carboxylase small subunit.

5. The nucleic acid construct of claim 1 wherein said N-terminal domain is an N-terminal domain of a mature maize ribulose-1,5-bisphosphate carboxylase small subunit.

6. The nucleic acid construct of claim 1 wherein said first or second transit peptide and said N-terminal domain are from the same 1,5-bisphosphate carboxylase small subunit protein.

7. The nucleic acid construct of claim 1 wherein said first or second transit peptide is a transit peptide of a sunflower ribulose-1,5-bisphosphate carboxylase small subunit.

8. The nucleic acid construct of claim 1 wherein said N-terminal domain is an N-terminal domain of mature sunflower ribulose-1,5-bisphosphate carboxylase small subunit.

9. The nucleic acid construct of any of claims 1, 5, 6 or 8 wherein said N-terminal domain comprises about 10 to about 40 amino acids.

10. The nucleic acid construct of any of claim 1, 5, 6 or 8 wherein said N-terminal domain comprises about 18 to about 33 amino acids.

11. A nucleic acid construct which codes for a polypeptide sufficient for localization of a gene product in a chloroplast of a plant cell which polypeptide comprises a fusion which in the direction of translation comprises a first chloroplast transit peptide from a sunflower ribulose-1,5-bisphosphate carboxylase small subunit, approximately 22 amino acids from the N-terminal region Of a mature maize ribulose-1,5-bisphosphate carboxylase small subunit and a second chloroplast transit peptide from a maize ribulose-1,5-bisphosphate carboxylase small subunit.