Ubiquitin regulatory system from maize

Patents owned by Mycogen Plant Science (probably currently controlled by Dow AgroSciences)

Patent number

Title, Summary of Claims and Independent Claims

Assignee

US 5510474

  • Earliest priority – 17 May 1988
  • Filed – 25 August 1994
  • Granted – 23 April 1996
  • Expected expiry – 23 April 2013
Title – Plant ubiquitin promoter system

Claim 1
An isolated DNA fragment, useful in effecting expression in both monocots and dicots of coding sequences placed 3′ to said fragment, wherein said DNA is approximately 2 kb in length, and said DNA fragment further comprises, in the following order beginning with the 5′ most element and proceeding toward the 3′ terminus of said DNA fragment:(a) two heat shock elements, which overlap;
(b) a promoter comprising a transcription start site;
(c) an intron of about 1 kb in length; and
(d) a translation start site; wherein said DNA fragment comprising said elements (a)-(d) regulates gene expression in both dicots and monocots, and wherein said DNA fragment comprises the nucleotide sequence shown from position -899 to 1092 of the maize ubiquitin sequence listed in FIG. 2.

Independent claim 1 is drawn to:

  • An isolated DNA fragment from -899 to +1092 of the 5′ flanking region of a maize ubiquitin sequence comprising from 5′ to 3′: two overlapping heat shock elements, a promoter with a transcription start site, an intron of about 1 kb, and a translation start site.
  • The DNA fragment has to be capable of regulating gene expression (of any gene) in both dicots and monocots; and
  • must include a portion of the sequence shown in Figure 2 of the patent (i.e., the sequence from position -899 to 1092).

Mycogen Plant Sciences

US 5614399

  • Earliest priority – 17 May 1988
  • Filed – 5 June 1995
  • Granted – 25 March 1997
  • Expected expiry -25 March  2014
Title – Plant ubiquitin promoter system

Claim 1

A method for selective heat shock induced enhancement of the constitutive expression of a structural gene in a plant cell comprising the steps of:
(a) transforming said plant cell with a DNA construct comprising an approximately 2 kb plant ubiquitin regulatory region operably joined to a DNA sequence of interest, wherein said plant ubiquitin regulatory region is from a plant ubiquitin gene and comprises at least one heat shock element, a promoter, a transcription start site, and an intron; and
(b) selectively applying stress conditions of high temperature to said transformed plant cell thereby inducing enhancement in expression of said DNA sequence of interest.

This patent is a divisional of US 5510474The claims are drawn to:

A method for rendering a constitutively expressed structural gene of a plant cell heat shock inducible by transforming the plant cell with a DNA construct which comprises a plant ubiquitin regulatory region and selectively applying high temperature to the transformed cell.

The plant ubiquitin regulatory region comprises:

  • at least one heat shock element
  • a promoter
  • a transcription start site
  • and an intron

The plant can be any type but the gene must be a structural gene (defined in the patent as one which encodes a protein, polypeptide or portion thereof) and so the claim does not cover constructs for expressing siRNAs.

The term “high temperature” is not defined but the patent describes that heat shock can occur at 40° C.

The patent discloses a single type of ubiquitin regulatory system – the sequence from a single maize ubiquitin gene – though the claims encompass ubiquitin regulatory systems from any kind of plant.

US 6020190

  • Earliest priority – 17 May 1988
  • Filed – 18 November 1996
  • Granted – 1 February 2000
  • Expected expiry – 17 May 2008
Title – Plant ubiquitin promoter system

Claim 1
A DNA construct comprising:(a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains a heat shock element and an intron, said intron being located at 3′ to said heat shock element, and
(b) a plant-expressible structural gene wherein said structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.

This patent is a divisional of US 5614399The claims generally relate to:

  • A DNA construct having a plant ubiquitin regulatory sequence no larger than 2 kb that contains an intron located 3′ of a heat shock element, and a plant-expressible structural gene under the control of the ubiquitin regulatory sequence.

Although the claim recites that the construct must contain a plant expressible gene this does not necessarily mean that the gene is from a plant.

However, the claim does require that the gene is a structural gene (e.g., must encode a protein, polypeptide or portion thereof).

The patent describes that placing a structural gene under the regulatory control of a promoter or a regulatory element “means positioning the structural gene such that the expression of the gene is controlled by these sequences. In general, promoters are found positioned 5′ (upstream) to the genes that they control.”

US 6054574

  • Earliest priority – 17 May 1988
  • Filed – 9 June 1998
  • Granted – 25 April 2000
  • Expected expiry – 17 May 2008
Title – Plant ubiquitin promoter system

Claim 1

A DNA fragment approximately 2 kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains a promoter comprising
a transcription start site,
one or more heat shock elements positioned 5′ to said transcription start site, and
an intron positioned 3′ to said transcription start site,
wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots such that the level of said constitutive gene expression in monocots is about one-third that obtained in said inducible gene expression in monocots.

Claim 9

A recombinant DNA construct comprising:

a. A DNA fragment approximately 2 kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said plant ubiquitin regulatory system contains a promoter comprising
a transcription start site,
one more heat shock elements positioned 5′ to said transcription start site,
a translational start site, and
an intron positioned 3′ to said transcription start site and 5′ to said translational start site, wherein said plant ubiquitin regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots such that said constitutive gene expression in monocots is at a level about one-third that obtained in said inducible gene expression in monocots, and

b. a plant-expressible heterologous structural gene positioned 3′ to said plant ubiquitin regulatory system and a polyadenylation signal positioned 3′ to said structural gene, wherein said heterologous gene is placed under the regulatory control of said plant ubiquitin regulatory system.

Claim 18

A DNA fragment, useful in effecting expression in both monocots and dicots of coding sequences placed 3′ to said fragment, wherein said DNA is isolated or incorporated into a larger piece of DNA but in a position other than in the 5′ sequence of a plant ubiquitin gene, is approximately 2 kb in length, and said DNA fragment further comprises, in the following order beginning with the 5′ most element and proceeding toward the 3′ terminus of said DNA fragment:

(a) one or more heat shock elements, which elements may or may not be overlapping;
(b) a promoter comprising a transcription start site; and
(c) an intron of about 1 kb in length; and wherein said DNA fragment comprising said elements (a)-(c) is capable of regulating gene expression in both dicots and monocots.

This patent is a divisional of US 6020190. The claims generally relate to:

  • A DNA fragment of approximately 2 kb comprising a plant ubiquitin regulatory system comprising:  one or more heat shock elements 5′ to a promoter with a transcription start site and an intron 3′ to the transcription start site. The system can be used to regulate constitutive and inducible expression for both dicots and monocots; however, constitutive expression must be 1/3 the level of heat-shock inducible expression in monocots.
  • A recombinant DNA construct having a plant ubiquitin regulatory system as described and a plant expressible heterologous structural gene under control of the regulatory region which also includes a polyadenylation sequence 3′ of the structural gene.
  • A DNA fragment of approximately 2kb as described above which can be part of a larger sequence so long as it is not 5′ of a plant ubiquitin gene. The level of heat-shock inducibility in monocots is not an element of this claim.

A particular level of heat shock induction in dicots is not an element of any of the claims.

The ubiquitin regulatory system can be from any plant although the patent does not describe any sequences other than those from maize.

CA 1339684

  • Earliest priority – 17 May 1988
  • Filed – 16 May 1989
  • Granted – 24 February 1998
  • Expected expiry – 24 February 2015
Title – Plant ubiquitin promoter system

Claim 1

A DNA fragment approximately 2kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:
a) overlapping heat shock elements and
b) an intron, and
wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots.

Claim 11
A recombinant DNA construct comprising:
a) a DNA fragment approximately 2kb in length, said DNA fragment comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:
1. overlapping heat shock elements and
2. an intron; and
wherein said regulatory system is capable of regulating constitutive and inducible gene expression in both dicots and monocots, and

b) a plant-expressible heterologous structural gene wherein said heterologous structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.

Claim 23

A method for the constitutive expression of a structural gene and the selected stress-induced enhancement in expression of said structural gene in a plant cell comprising the steps of:
a) transforming said plant cell with a DNA construct comprising an approximately 2kb plant ubiquitin regulatory system, wherein is found a heat shock element and an intron, and
b) a plant-expressible structural gene that is under the regulatory control of said plant regulatory system, and
c) selectively applying stress conditions of high temperature to said transformed plant cell thereby inducing enhancement in expression of said structural gene.

The claims are generally drawn to:

  •  plant ubiquitin regulatory system of around 2 kb capable of constitutive and inducible regulation of gene expression in both monocots and dicots. The system contains an overlapping heat shock elements and intron.
  • A recombinant DNA construct having a plant ubiquitin regulatory system as described and a plant expressible structural gene under control of the regulatory region.
  •  A method for both constitutive and stress-inducible enhanced expression of a structural gene in a plant cell. The transformed cell with the plant ubiquitin system is subjected to high temperature to induce enhancement in gene expression.

The construct used in method claim 23 does not require overlapping heat shock elements but encompasses constructs with a single heat shock element.

The claims don’t require a particular level of induction after heat shock.

As with the above patents, the ubiquitin regulatory system could be from any plant so long as that system comprises a heat shock element.

EP 342926 B1

  • Earliest priority – 17 May 1988
  • Filed – 16 May 1989
  • Granted – 28 September 1994
  • Expected expiry – 16 May 2009
Title – Plant ubiquitin promoter system

Claim 1
A DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:
a) a heat shock element and
b) an intron.
Claim 10
A DNA construct comprising:a) a DNA sequence no larger than 2 kb, said DNA sequence comprising a plant ubiquitin regulatory system, wherein said regulatory system contains:
1. a heat shock element, and
2. an intron; and

b) a plant-expressible structural gene wherein said structural gene is placed under the regulatory control of said plant ubiquitin regulatory system.

Claim 12
A method for the constitutive expression of a structural gene and the selected stress-induced enhancement in expression of said structural gene in a plant cell comprising the steps of:a) transforming said plant cell with a DNA construct comprising:
i. a plant ubiquitin regulatory system, wherein is found a heat shock element and an intron, and
ii. a plant-expressible structural gene that is under the regulatory control of said plant ubiquitin regulatory system, and

b) selectively applying stress conditions to said transformed plant cell thereby inducing enhancement in expression of said structural gene.

Remarks

The European patent converted to and in force in Belgium (BE), France (FR), Germany (DE), Great Britain (GB), Italy (IT), Spain (ES) and the Netherlands (NL). A patent (JP 3509855) was also granted in Japan.

Note: Patent information was last updated on 23 May 2006. Search terms: “ubiquitin” in abstract and “Mycogen” in applicant. Patent database: PatentLens in combination with INPADOC.

Patents and applications relating to the ubiquitin regulatory region owned by Monsanto

Patent/application number

Title, Independent Claims and Summary of Claims

Assignee

US 6878818

  • Earliest priority – 9 September 1999
  • Filed – 12 June 2000
  • Granted – 12 April 2005
  • Expected expiry – 9 September 2019
Title – Modified ubiquitin regulatory system

Claim 1

An isolated DNA sequence comprising a ubiquitin regulatory system lacking heat shock elements wherein the ubiquitin regulatory systemcomprises the nucleotide sequence according to SEQ.ID.NO. 8.

The claims are to:

  • DNA sequences which comprise a specific nucleotide sequence (SEQ.ID.NO. 8), a 2033 bp sequence which includes the promoter, the 83 bp untranslated first exon and the intron of the maize Ubi-1 gene.
  • Dependent claims are drawn to host cells, transgenic plants (of any kind), and seeds comprising the sequences,  as well as constructs comprising the sequence operably linked to a plant-expressible structural coding sequence and vectors comprising the constructs.

Claim 1 requires that the DNA sequence has at least the 2033 bp sequence but allows for the inclusion of additional sequences – such as the remainder of the coding region of the Ubi-1 gene or heterologous sequences not usually found in association with the Ubi-1 promoter and intron. However, the heat shock elements cannot be included.

Because the claims do require SEQ.ID.NO. 8 as an element of the sequence, a regulatory system which included portions of the promoter and intron sequence required for expression, but which deleted other parts of the sequence would not literally be covered by these claims.

Monsanto UK Ltd

AU 769567 B2

  • Earliest priority – 9 September 1999
  • Filed – 7 September 2000
  • Granted – 29 January 2004
  • Expected expiry – 7 September 2020
Title – Modified ubiquitin regulatory system

Claim 1
A DNA sequence comprising a ubiquitin regulatory system lacking heat shock elements wherein the ubiquitin regulatory system comprisesthe nucleotide sequence according to SEQ.ID.NO. 8.

Claim 1 of the Australian patent is identical to that of the US patent.

EP 1210446 A1

  • Earliest priority – 9 September 1999
  • Filed – 7 September 2000
  • Status – Pending
  • Expected expiry – N/A
Title – Modified ubiquitin regulatory system

Claim 1

A DNA sequence comprising an ubiquitin regulatory system lacking heatshock elements.

Claim 2

A DNA sequence comprising an ubiquitin regulatory system lacking heatshock elements.

These independent claims are broader than those in the US patent since the exact base composition of the ubiquitin regulatory system is not an element of the claims and the system can be from any organism.

The application discloses that a ubiquitin regulatory system which lacks heat shock elements includes a promoter, a transcription start site, an intron and optionally a translation start site. Only the regulatory sequences of one maize ubiquitine gene is disclosed.

CA 2384517

  • Earliest priority – 9 September 1999
  • Filed – 7 September 2000
  • Status- Pending
  • Expected expiry – N/A
Title – Modified ubiquitin regulatory system

Claim 1
A DNA sequence comprising a ubiquitin regulatory system lacking heat shock elements wherein the ubiquitin regulatory system comprisesthe nucleotide sequence according to SEQ.ID.NO. 8.

The claims of this Canadian application are exactly the same as  AU 769567 B2.

Note: Patent information was last updated on 23 May 2006. Search terms: “ubiquitin” in abstract and “Monsanto” in applicant. Patent database: PatentLens and esp@cenet in combination with INPADOC.

Patent owned by Prodigene

Patent/application number

Title, Independent Claims and Summary of Claims

Assignee

US 6977325

  • Earliest priority – 9 Jun 2000
  • Filed -28 Feb 2002
  • Granted- 20 Dec 2005
  • Expected expiry – 16 Jun 2020 (adjusted)

Title – Plant promoter sequences and methods of use for same

Claim 1
An engineered ubiquitin promoter sequence capable of directing expression of a nucleotide sequence in a plant cell, said engineered ubiquitin promoter sequence comprising: a heat shock region, wherein said heat shock region has the sequence as set forth in SEQ ID NO: 4.
Claim 2
A method for causing expression of a heterologous structural gene or open reading frame in a plant cell, said method comprising: introducing to a plant cell an expression construct comprising an engineered ubiquitin promoter sequence operably linked to said heterologous structural gene or open reading frame, wherein said engineered ubiquitin promoter sequence comprises a heat shock region, wherein said heat shock region has the sequence as set forth in SEQ ID NO: 4.

Prodigene Inc.

Remarks

The patent application filed in Australia (AU 200175433) lapsed. The related PCT application is WO 2001/094394.

Note: Patent information was last updated on 30 October 2006. Search terms: “ubiquitin” in abstract and “Prodigene” in applicant. Patent database: PatentLens and in combination with INPADOC.