Enhanced expression in plants using non-translated leader sequences

• US 5,362,865 A
• Filed: 09/02/1993
• Issued: 11/08/1994
• Est. Priority Date: 09/02/1993
• Status: Expired due to Term

• Current Assignee: Monsanto Technology LLC (Bayer AG)
• Original Assignee: Monsanto Company (Bayer AG)
• Inventors: Austin, Glenn D.
• Primary Examiner(s): Fox, David T.
• Application Number:
  US08/117,374
• Time in Patent Office: 432 Days
• Field of Search: 536/24.1, 536/24.5, 435/172.3, 435/240.4, 800/205
• US Class Current: 536/24.1
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells

Abstract

This invention provides DNA molecules which comprise 5'"'"' non-translated leader sequences derived from genes coding for heat shock proteins that enhance gene expression in plants when present in a chimeric gene. Plant cells and plants containing same are also provided herewith. Further provided is a method for enhancing gene expression in plants.

Claims

1. A DNA molecule which comprises:

(a) a promoter region which functions in plant cells to cause the production of an RNA sequence;

which is operably linked to

(b) a non-translated leader sequence derived from a plant heat shock gene selected from the group consisting of petunia HSP70, soybean HSP17.9, and maize HSP70, wherein said non-translated leader sequence is heterologous to said promoter;

which is operably linked to

(c) a structural DNA sequence, wherein said structural DNA sequence is heterologous to said non-translated leader sequence;

which is operably linked to

(d) a 3'"'"' non-translated sequence that functions in plant cells to cause the termination of transcription and the addition of polyadenylated ribonucleotides to the 3'"'"' end of the transcribed mRNA sequence.

2. A DNA molecule according to claim 1 wherein said promoter region is selected from the group consisting of an FMV35S promoter region, a CaMV35S promoter region, and an enhanced CaMV35S promoter region.

3. A DNA molecule according to claim 1 wherein said structural DNA sequence is in the antisense orientation.

4. A DNA molecule according to claim 1 wherein said 3'"'"' untranslated region is from a gene selected from the group consisting of the nopaline synthase (NOS) gene, the soybean 7S storage protein genes, and the pea small subunit of the ribulose 1,5-bisphosphate carboxylase-oxygenase (ssRUBISCO) E9 gene.

5. A method for providing enhanced gene expression in plants which comprises:

(a) a transforming plant cells with a DNA molecule which comprises;

(i) a promoter region which functions in plant cells to cause the production of an RNA sequence;

which is operably linked to

(ii) a non-translated leader sequence derived from a plant heat shock gene selected from the group consisting of petunia HSP70, soybean HSP17.9, and maize HSP70, wherein said non-translated leader sequence is heterologous to said promoter;

which is operably linked to

(iii) a structural DNA sequence, wherein said structural DNA sequence is heterologous to said non-translated leader sequence;

which is operably linked to

(iv) a 3'"'"' non-translated DNA sequence which functions in plant cells to cause the termination of transcription and the addition of polyadenylated ribonucleotides to the 3'"'"' end of the transcribed mRNA sequence;

(b) selecting said plant cells which have been transformed;

(c) regenerating said plant cells to provide a differentiated plant; and

(d) selecting a transformed plant which expresses said structural gene.

6. A method according to claim 5 wherein said promoter region is selected from the group consisting of an FMV35S promoter region, a CaMV35S promoter region, and an enhanced CaMV35S promoter region.

7. A method according to claim 5 wherein said structural gene is in the antisense orientation.

8. A method according to claim 5 wherein said 3'"'"' untranslated region is from a gene selected from the group consisting of the nopaline synthase (NOS) gene, the soybean 7S storage protein genes, and the pea small subunit of the ribulose, 1,5-bisphosphate carboxylase-oxygenase (ssRUBISCO) E9 gene.

9. A transformed plant cell containing a DNA molecule which comprises:

(a) a promoter region which functions in plant cells to cause the production of an RNA sequence;

which is operably linked to

(b) a non-translated leader sequence derived from a plant heat shock gene selected from the group consisting of petunia HSP70, soybean HSP17.9, and maize HSP70, wherein said non-translated leader sequence is heterologous to said promoter;

which is operably linked to

(c) a structural DNA sequence, wherein said structural DNA sequence is heterologous to said non-translated leader sequence;

which is operably linked to

(d) a 3'"'"' non-translated sequence that functions in plant cells to cause the termination of transcription and the addition of polyadenylated ribonucleotides to the 3'"'"' end of the transcribed mRNA sequence.

10. A plant cell according to claim 9 wherein said promoter region is selected from the group consisting of an FMV35S promoter region, a CaMV35S promoter region, and an enhanced CaMV35S promoter region.

11. A plant cell according to claim 9 wherein said structural gene is in the antisense orientation.

12. A plant cell according to claim 9 wherein said 3'"'"' untranslated region is from a gene selected from the group consisting of the nopaline synthase (NOS) gene, the soybean 7S storage protein genes, and the pea small subunit of the ribulose 1,5-bisphosphate carboxylase-oxygenase (ssRUBISCO) E9 gene.