Fruit Promoters

Summary

Among the large patent portfolio of Calgene on tissue-specific promoters, there are three main patent families containing granted patents directed to fruit-specific regulatory regions. Patents and patent applications that were assigned to Calgene may now be held by Monsanto, to which any inquiries about licensing should be directed (http://www.monsanto.com/monsanto/layout/about_us/contactus.asp).

Patents of one of the patent families are drawn to DNA constructs containing promoters “preferentially expressed in fruit tissues”.  Although the basic patents in the patent family are involved in the expression of genes regulating fruit ripening and drive the expression of genes of interest in mature ovaries, the definitions are broad, and examples in which seed-specific expression is cited suggest that tissues such as seed, fruit integument, cotton fibers and so on would be construed as “fruit tissues”. In addition, general methods to regulate the fruit phenotype are part of the patented inventions.

In the other two patent families, the definitions are more specific.  One relates primarily to cotton fiber production, although it contains some broader claims. In the second, the promoters of the invention are expressed in receptacle tissue, a flower part that makes most of the fleshy tissue in accessory fruits such as strawberry, apple and pear. The genes driven by the promoter influence fruit development, maturation and ripening.  Some analysis of this patent family is provided below.

Broadest patent family

The claims of the United States patent are drawn to methods for altering the phenotype of the fruit tissue of a plant transformed with DNA constructs comprising:

  • a promoter preferentially expressed in a fruit tissue (note that this does not mean “fruit-specific”);
  • a DNA sequence of interest different from the native gene of the promoter (note that this need not be a “gene”–could be antisense or RNAi); and
  • a transcriptional termination region.

Unlike the European patent, the genes from which the fruit-specific promoters are obtained are not limited to anthesis process or fruit maturation and ripening. The promoters are from genes preferentially transcribed in fruit tissue. Because there is no limitation in the fruit gene promoter used in the DNA construct to alter the phenotype of a fruit, the invention claimed in the United States patent, although directed to methods, is broader in scope than in the Canadian and European patents. Also, the claimed methods are described in very broad terms.

Calgene’s European patent claims:

  • A DNA construct comprising a fruit transcriptional initiation region from a gene that becomes active during anthesis (period during which a flower is fully open and functional), remains active until the ripe period and is transcribed in mature ovary tissue. The construct includes a gene of interest under the control of such transcriptional region and a transcriptional termination region.
  • A DNA construct having a fruit promoter of a plant storage protein that becomes and remains active during the stages described above.
  • A method for the modification of the phenotype of a tomato fruit. The sort of phenotypic modification is not specified in the claim.
  • A tomato cell is transformed with a DNA construct as described above. A tomato plant is regenerated and grown from the transformed cell.
Patent Number Title, Independent Claims and Summary of Claims Assignee
US 4943674

  • Earliest priority -17 January 1985
  • Filed – 26 May 1987
  • Granted – 24 July 1990
  • Expected expiry – 24 July 2007
Title – Fruit specific transcriptional factors

Claim 1
A DNA construct comprising in the direction of transcription,
a tomato 2All transcriptional initiation region joined to a DNA sequence of interest, wherein said DNA sequence of interest is other than the wild-type sequence and is under the transcriptional regulation of said 2All initiation region and
a transcriptional termination region.
Claim 5
A DNA construct comprising in the direction of transcription,
a tomato 2All transcriptional initiation region joined to a DNA sequence of interest, wherein said DNA sequence is other than the wild-type sequence and comprises a unique restriction site for insertion of a second DNA sequence of interest to be under the transcriptional regulation of said 2All initiation region, and
a transcriptional termination region.
Claim 9
A method for modifying the phenotype of tomato fruit said method comprising:
transforming a tomato plant cell with a DNA construct under genomic integration conditions, wherein said DNA construct comprises in the direction of transcription,
a tomato 2All transcriptional initiation region jointed to a DNA sequence other than the wild-type sequence and capable of modifying the phenotype of fruit cells upon transcription, wherein said DNA sequence is under the transcriptional regulation of said initiation region and
a transcriptional termination region, whereby said DNA construct becomes integrated into the genome of said tomato plant cell;         regenerating a plant from said transformed tomato plant cell; and
growing said plant to produce tomato fruit of the modified phenotype.
Claim 12
Tomato fruit comprising a DNA construct comprising in the direction of transcription,
a tomato 2All transcriptional initiation region joined to a DNA sequence of interest, wherein said DNA sequence is other than the wild-type sequence and is under the transcriptional regulation of said 2All initiation region, and
a transcriptional termination region.
Claim 13
A tomato plant comprising: a DNA construct comprising in the direction of transcription,
a tomato 2All transcriptional initiation region joined to a DNA sequence of interest, wherein said DNA sequence is other than the wild-type sequence and is under the transcriptional regulation of said 2All initiation region, and
a transciptional termination region.
Claim 14
A tomato plant comprising: a DNA construct comprising in the direction of transcription,
a tomato 2All transcriptional initiation region joined to a DNA sequence of interest, wherein said DNA sequence is other than the wild-type sequence and comprises a unique restriction site for insertion of a second DNA sequence of interest to be under the transcriptional regulation of said 2All initiation region, and
a transciptional termination region.

Calgene
Inc.

US 5753475

  • Earliest priority -17 January 1985
  • Filed – 10 August 1993
  • Granted – 19 May 1998
  • Expected expiry – 19 May 2015
Title – Methods and compositions for regulated transcription and expression of heterologous genes

Claim 1
A method for obtaining a plant having a regulatable phenotype, said method comprising:
transforming a host plant cell with a DNA construct under genomic integration conditions, wherein said construct comprises as operably linked components in the direction of transcription,
a promoter region obtainable from a gene, wherein transcription of said gene is preferentially regulated in a plant fruit tissue,
a DNA sequence of interest other than the native coding sequence of said gene, and
a transcription termination region, wherein said components are functional in a plant cell, whereby said DNA construct becomes integrated into a genome of said plant cell;
regenerating a plant from said transformed plant cell, and
growing said plant under conditions whereby said DNA sequence of interest is expressed and a plant having said regulatable phenotype is obtained.
Claim 2
A method for altering the phenotype of fruit tissue as distinct from other plant tissue, said method comprising:
growing a plant, wherein said plant comprises cells containing a DNA construct integrated into their genome, said DNA constructcomprising, in the 5′ to 3′ direction of transcription,
a transcriptional initiation region from a gene, wherein transcription of said gene is preferentially regulated in a plant fruit tissue,
a DNA sequence of interest other than the coding sequence native to said transcriptional initiation region, and
a transcriptional termination region, whereby said DNA sequence of interest is transcribed under transcriptional control of said transcriptional initiation region and a plant having an altered phenotype is obtained.
Claim 5
A method for modifying the genotype of a plant to impart a desired characteristic to fruit as distinct from other plant tissue, said method comprising:
transforming under genomic integration conditions, a host plant cell with a DNA construct comprising in the 5′ to 3′ direction of transcription,
a transcriptional initiation region from a gene, wherein transcription of said gene is preferentially regulated in a plant fruit tissue,
a DNA sequence of interest other than the native coding sequence of said gene, and
a transcriptional termination region, whereby said DNA construct becomes integrated into the genome of said plant cell;
regenerating a plant from said transformed host cell; and
growing said plant to produce fruit having a modified genotype.
Claim 9
A method for modifying transcription in fruit tissue as distinct from other plant tissue, said method comprising:
growing a plant capable of developing fruit tissue under conditions to produce fruit, wherein said plant comprises cells containing a DNA construct integrated into their genome, said DNA construct comprising, in the 5′ to 3′ direction of transcription,
a fruit-specific transcriptional initiation region,
a DNA sequence of interest other than the coding sequence native to said transcriptional initiation region, and
a transcriptional termination region, whereby said DNA sequence of interest is transcribed under transcriptional control of said fruit-specific transcription initiation region.
Claim 12
A method to selectively express a heterologous DNA sequence of interest in fruit tissue as distinct from other plant tissue, said method comprising:growing a plant capable of developing fruit tissue under conditions to produce fruit, wherein said plant comprises cells having a genomically integrated DNA construct comprising, as operably linked components in the 5′ to 3′ direction of transcription,
a fruit-specific transcriptional initiation region and a translational initiation region,
a DNA sequence of interest other than the coding sequence native to said transcriptional initiation region,
a transcriptional termination region downstream of said DNA sequence of interest, whereby said DNA sequence of interest is expressed under control of said fruit-specific transcriptional and translational initiation region.

This patent is also a Continuation in part of US 5420034 (see the Seed promoter section).

EP 316441 B1

  • Earliest priority – 26 May 1987
  • Filed – 26 May 1988
  • Granted – 5 December 2001
  • Expected expiry – 26 May 2008
Title – Fruit-specific transcriptional factors

Claim 1
A DNA construct comprising in the direction of transcription,
a fruit-specific transcriptional initiation region from a gene which becomes active at or immediately after anthesis and remains active at least until the ripe period, and which is transcribed in mature ovary tissue, joined to a DNA sequence of interest other than the wild-type sequence associated with said initiation region, wherein said DNA sequence of interest is under the transcriptional regulation of said initiation region anda transcriptional termination region.
Claim 7
A DNA construct comprising in the direction of transcription,
a fruit-specific transcriptional initiation region of a plant storage protein which becomes active at or immediately after anthesis and remains active at least until the ripe period and which is transcribed in mature ovary tissue, joined to a DNA sequence other than the wild-type sequence, wherein said sequence comprises a unique restriction site for insertion of a sequence of interest to be under the transcriptional regulation of said initiation region, and
a transcriptional termination region.
Claim 12
A method for specifically modifying the phenotype of fruit substantially distinct from other plant tissue, said method comprising:
transforming a plant cell with a DNA construct under genomic integration conditions, wherein said DNA construct comprises in the direction of transcription,
a fruit-specific transcriptional initiation region which becomes active at or immediately after anthesis, and remains active at least until the ripe period and which is transcribed preferentially in mature ovary tissue, joined to a DNA sequence other than the wild-type sequence and capable of modfying the phenotype of fruit cells upon transcription, wherein said sequence is under the transcriptional regulation of said initiation region, and
a transcriptional termination region, whereby said DNA construct becomes integrated into the genome of said plant cell;
regenerating a plant from said transformed plant cell; and
growing said plant to produce fruit of the modified phenotype.

Remarks

A related Canadian application (CA 1338827) has lapsed and a related patent was granted in New Zealand (NZ 224787).  There is also a related patent in Australia. Related patent applications also filed in Israel (IL 86515 A0), and Japan (JP 2500163 T2). Related patent application in China (CN 1036305 A) was withdrawn 20 March 1991.

Note: Patent information was last updated on 15 May 2006. Search terms: “promoter” in abstract and “Calgene” in applicant. Patent database: PatentLens in combination with INPADOC.

Cotton fiber patent family

As mentioned above, Cotton (Gossypium hirsutum) fiber can be construed as a type of “fruit tissue” in broader botanical terms. Calgene has a United States patent  directed to a promoter from cotton expansin gene. The cotton expansin gene is expressed in developing fiber and, according to the specification, the promoter of the cotton expansin gene can be used to drive a gene of interest in developing cotton fiber for modifying cotton fiber phenotypes.

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 6566586

  • Earliest priority – 7 January 1997
  • Filed – 7 January 1998
  • Granted – 20 May 2003
  • Expected expiry – 7 January 2018
Title – Cotton expansin promoter sequence

Claim 1
An isolated DNA sequence comprising the sequence of SEQ ID NO: 1.

The only independent claim is drawn to an isolated DNA sequence comprising the 2614 bp cotton expansin promoter sequence. A recombinant DNA construct comprising the promoter sequence and a plant comprising a plant cell comprising the DNA construct are also claimed in the dependent claims.

Calgene LLC

Remarks

Related European application (EP 968292 A1) was withdraw and the application in Australia (AU 57322/98) also lapsed.

Note: Patent information was last updated on 18 May 2006. Search terms: “promoter” in abstract and “Calgene” in applicant. Patent database: PatentLens in combination with INPADOC.

Receptacle patent family

A European patent application and a granted United States patent filed by Calgene are directed to fruit promoters in general driving the expression of genes in the receptacle tissue of a fruit.

Specific Patent Information

Patent Number Title, Summary of Claims and Independent Claims Assignee
EP 973922

  • Earliest priority- 6 February 1998
  • Filed – 4 February 1999
  • Withdrawn – 7 January 2004
Title – Strawberry fruit promoters for gene expression

The claims as filed of the European patent application are broader in this case, and recite:

  • DNA constructs comprising a transcriptional factor driving the expression of a heterologous gene in the receptacle tissue of a fruit. The expression of the gene of interest is either:
    • increased during fruit ripening or
    • decreased during fruit development and maturation.

Calgene Inc.

US 6043410

  • Earliest priority- 6 February 1998
  • Filed – 6 February 1998
  • Granted – 28 March 2000
  • Expected expiry – 6 February 2018
Title – Strawberry fruit promoters for gene expression

Claim 1
A DNA construct comprising the promoter sequence from RJ39, isolated from Fragaria, operably-linked to a heterologous DNA coding sequence of interest.
CA 2285465

  • Filed – 4 February 1999
  • Expiration – Dead application 4 February 2004
Title – Strawberry fruit promoters for gene expression

Dead application

Note: Patent information was last updated on 15 May 2006. Search terms: “promoter” in abstract and “Calgene” in applicant. Patent database: PatentLens in combination with INPADOC.