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Cold-inducible promoters

Promoters responsive to low or cold temperatures (equal to or below 10°C) are the subject matter of patents and patent applications filed by the Japanese companies Takara Shuzo Co., Ltd and Japan Tobacco Inc., The University of Quebec in Montreal and Danisco.

Most of the cited institutes claim in their patents a particular “cold” promoter and its variant sequences.

A. Takara Shuzo Co., Ltd’s patents and patent applications

The United States granted patents and the European and Canadian patent applications relate to a vector useful for the expression of a protein under low temperature conditions. The vector of the invention contains:

  1. a promoter regulatory region; and
  2. a 5′-untranslated region derived from a cold-shock protein gene.

The promoter region and 5′ untranslated region of the vector are not limited to a particular promoter. In addition a promoter and a 5′-untranslated region are derived from the cold-shock protein (csp) A of Escherichia coli. The nucleotide sequence of such promoter is also claimed by the inventors.

Patent Number Title, Independent Claims and Summary of Claims Assignee
US 6479260

  • Earliest priority – 20 November 1997
  • Filed – 19 May 2000
  • Granted – 12 November 2002
  • Expected expiry – 19 May 2020
 Title – Low-temperature inducible expression vector

Claim 1
A vector which is characterized in containing each of the following elements:
(1) a promoter which shows its action in the host to be used;
(2) regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5′-untranslated region derived from cold-shock protein gene mRNA or a region which codes for the region where substitution, deletion, insertion or addition of at least one base is applied to the said untranslated region and which contains a base sequence as shown in SEQ ID NO:1 in the Sequence Listing.
Claim 10
An isolated promoter consisting of a base sequence as shown in SEQ ID NO: 5 in the Sequence Listing.
Claim 11
An isolated promoter containing a base sequence as shown in SEQ ID NO: 5 in the Sequence Listing and consisting of a base sequence having 135 or less bases, wherein the promoter does not contain the region which is ascribed to mRNA.

Takara Shuzo Co., Ltd

(now Takara Holdings Inc.)
US 6897042

  • Earliest priority – 20 November 1997
  • Filed – 10 October 2002
  • Granted – 24 May 2005
  • Expected expiry – 19 May 2020
 Title – Low-temperature inducible expression vector

Claim 1
A vector which is characterized in containing each of the following elements:
(1) a promoter which is derived from cold-shock protein gene, and which shows its action in the host to be used;
(2) regulatory region for regulating the action of the promoter of (1), wherein said regulatory region is located downstream of the promoter of (1); and
(3) a region which codes for the 5′-untranslated region derived from cold-shock protein gene mRNA or a region which codes for the region where substitution, deletion, insertion, or addition of at least one base is applied to said untranslated region.
Claim 12
A method for the expression of the desired protein which is characterized in containing the following steps:
1) a step where a host is transformed with a vector containing:
a) a promoter which is derived from cold-shock protein gene and which shows its action in the host to be used;
b) a regulatory region for regulating the action of the promoter of a), wherein said regulatory region; and
c) a region which codes for the 5′-untranslated region derived from cold-shock protein gene mRNA or a region which codes for the region where substitution, deletion, insertion, or addition of at least one base is applied to the untranslated region wherein a gene coding for the desired protein is integrated;
2) a step where the resulting transformant is incubated; and
3) a step where action of promoter is induced via a function of a regulatory region and, at the same time, incubating temperature is made lower than the ordinary temperature to express the desired protein.

This patent is a Countinuation of US 6479260.
EP 1033408 A1

  • Earliest priority – 20 November 1997
  • Filed – 17 November 1998
  • Granted –  Pending
  • Expected expiry – N/A
 Title – Cold-inducible expression vector

Claim 1
A vector which is characterized in containing each of the following elements:(1) a promoter which shows its action in the host to be used;
(2) regulatory region for regulating the action of the promoter of (1); and
(3) a region which codes for the 5′-untranslated region derived from cold-shock protein gene mRNA or a region which codes for the region where substitution, deletion, insertion or addition of at least one base is applied to the said untranslated region.
Claim 14
A promoter containing a base sequence as shown in SEQ ID NO:5 in the Sequence Listing and consisting of a base sequence having 135 or less bases.
CA 2309600 AA

  • Earliest priority – 20 November 1997
  • Filed – 17 November 1998
  • Granted –  Pending
  • Expected expiry – N/A
 Title – Cold-inducible expression vector

The claims of this Canadian patent application are the same as EP 1033408 (above).

Remarks

 The related Australian application AU 10546/99 lapsed on August 3, 2000.

Note: Patent information was last updated on 10 May 2006.

Search terms: “promoter” in abstract and “Takara Shuzo” in applicant.

Patent database: PatentLens.

B. Patents on particular cold-temperature promoters

As mentioned above, the patents filed by most institutes relate to a specific low temperature-responsive promoter.

The following table summarizes some bibliographic data of the patents and patent applications filed by the cited entities and a brief description of the promoter claimed.

Patent number Title, Independent Claims and Summary of Claims Assignee
US 6084089

  • Earliest priority – 27 December 1995
  • Filed –  27 October 1997
  • Granted – 4 July 2000
  • Expected expiry – 27 October 2017
 Title – Cold-inducible promoter sequences

Claim 1
A DNA sequence comprising a nucleotide sequence from the first to the 3546th nucleotide in the nucleotide sequence shown in SEQ ID NO:1.
Claim 2
A DNA sequence comprising a nucleotide sequence from the 2418th to the 3541st nucleotide in the nucleotide sequence shown in SEQ ID NO:1.
Claim 3
A DNA sequence comprising a nucleotide sequence from the first to the 4120th nucleotide in the nucleotide sequence shown in SEQ ID NO:2.

DNA sequences of a promoter isolated from potato. Partial and modified sequences of the promoter with cold-inducible promoter activity are also claimed.

According to the inventors, “cold-inducible” promoter means induced at a temperature not higher than 6°C and expression of the gene controlled by such promoter maintained for not less than 5 months.

Japan Tobacco
AU 725727 B2

  • Earliest priority – 27 December 1995
  • Filed – 26 December 1996
  • Granted – 19 October 2000
  • Expected expiry – 26 December 1996
 Title– Cold-inducible promoter sequences

Claim 1
A DNA sequence having a nucleotide sequence from first to 3546th nucleotide in the nucleotide sequence shown in SEQ ID. No:1, or a part thereof having a cold-inducible promoter activity, or a DNA sequence having the same nucleotide sequence as said DNA sequences except that one or more nucileotides are deleted or substituted, or one or more nucleotides are inserted or added, which DNA sequence has a cold-inducible promoter activity.
Claim 3
A DNA sequence having a nucleotide sequence from 2418th to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No:1, or a part thereof having a cold-inducible promoter activity, or a DNA sequence having the same nucleotide sequence as the said DNA sequences except that one or more nucleotides are deleted or substituted, or one or more nucleotides are inserted or added, which DNA sequence has a cold-inducible promoter activity.
Claim 4
A cold-inducible promoter sequence having a nucleotide sequence from 2418th to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No:1, or a DNA sequence having the same nucleotide sequence as said DNA sequence except that one or more nucleotides are deleted or substituted, or one or more nucleotides are inserted or added, which DNA sequence has a cold-inducible promoter activity.
Claim 5
A DNA sequence having a nucleotide sequence from first to 4 120th nucleotide in the nucleotide sequence shown in SEQ ID. NO:2, or a part thereof having a cold-inducible promoter activity, or a DNA sequence having the same nucleotide sequence as the said DNA sequences except that one or more nucleotides are deleted or substituted, or one or more nucleotides are inserted or added, which DNA sequence has a cold-inducible promoter activity.
Claim 7
A probe comprising a DNA fragment having at least 18 consecutive nucleotides  in the region  from  45th  to  839th nucleotide  in  the  sequence  shown  in  SEQ  ID  NO:3  in  the Sequence Listing or a sequence complementary thereto.
EP 812917 B1

  • Earliest priority -27 December 1995
  • Filed – 26 December 1996
  • Granted – 12 May 2005
  • Expected expiry – 26 December 2016
 Title – Cold-inducible promoter sequences

Claim 1
A DNA sequence having a nucleotide sequence from first to 3546th nucleotide in the nucleotide sequence shown in SEQ ID. No. 1, or a part thereof having a cold-inducible promoter activity.
Claim 2
A DNA sequence having a nucleotide sequence from 2418th to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No. 1, or a part thereof having a cold-inducible promoter activity.
Claim 3
A cold-inducible promoter sequence having a nucleotide sequence from 2418th to 3541st nucleotide in the nucleotide sequence shown in SEQ ID. No. 1.
Claim 4
A DNA sequence having a nucleotide sequence from first to 4120th nucleotide in the nucleotide sequence shown in SEQ ID. No. 2, or a part thereof having a cold-inducible promoter activity.
Claim 5
Use of a DNA fragment having at least 15 consecutive nucleotides in the region from 45th to 839th nucleotide in the sequence shown in SEQ ID. No. 3 in the Sequence Listing or a sequence complementary thereto, for the preparation of a probe for screening cold-inducible promoters.

Remarks

 The related patents were also granted in Japan (JP 3469902), China (CN 96192162.5) and Canada (CA 2213991).
US 6184443

  • Earliest priority – 21 October 1994
  • Filed – 15 September 1997
  • Granted – 6 February 2001
  • Expected expiry – 14 September 2017
 Title – Cold-inducible promoter and tuber-specific promoter sequence from potato alpha amylase gene

Claim 1
An isolated alpha-amylase promoter having cold-sensitive promoter activity, having a sequence comprising the 5.5 Kb EcoRI DNA fragment of Solanum tuberosum from the transformed E. coli strain, DH5alpha-gPAmy 351 (NCIMB Accession Number 40682).
Claim 12
An isolated alpha-amylase promoter having tuber-specific activity and having a sequence comprising SEQ ID NO: 1.

DNA sequences (variant, homologue and fragments) of an isolated alpha-amylase promoter from potato (Solanum tuberosum) having cold-sensitive activity. The promoter has tuber tissue specific activity.

Danisco

Remarks

 Related patent applications in Europe (EP 787194 A1) and Canada (CA 2202896 AA) were withdrawn and Lapsed, respectively. Related Australian (AU 27881/95) application has also lapsed.
US 5847102

  • Earliest priority – 12 April 1995
  • Filed – 12 April 1995
  • Granted – 8 December 1998
  • Expected expiry – 8 December 2015
 Title – Cold-induced promoter from winter Brassica napus

Claim 1
An isolated DNA molecule comprising a 5′ regulatory region of a low temperature-responsive gene BN115 from Brassica napus, comprising nucleotides 961-1210 of SEQ ID NO:1.
Claim 7
An isolated cold inducible promoter, or a functional fragment thereof capable of regulating the expression of a gene in response to a change in temperature, said cold inducible promoter comprising nucleotides 1-1271 of SEQ ID NO:1.
Claim 8
An isolated DNA molecule comprising a sequence of at least 15 contiguous nucleotides of a 5′ regulatory region of a low temperature responsive gene BN115 from Brassica napus, as defined by nucleotides 1-1271 of SEO ID NO:1.
Claim 13
An isolated enhancer, or a functional fragment thereof each capable of mediating the expression of a gene under the control of a promoter and said enhancer or fragment, said enhancer obtained from the regulatory region of BN115 from Brassica napus.
Claim 16
An isolated negative regulatory element capable of repressing gene expression at 22° C., obtained from the regulatory region of BN115 from Brassica napus and comprising nucleotides 461-623 of SEQ ID NO:1.

Isolated DNA sequences comprising a promoter and 5′ regulatory region of the low temperature responsive gene BN115 from Brassica napus. Sequences that hybridize to the promoter sequence and exhibit low temperature activity are also part of the claimed invention. An enhancer and a negative regulatory element present in the regulatory region are also claimed.

Canadian Ministry of Agriculture
CA 2146712

  • Earliest priority – 10 April 1995
  • Filed – 10 April 1995
  • Granted – 25 June 2002
  • Expected expiry – 10 April 2015
 Title – Cold-induced promoter from winter Brassica napus

The claims are the same as US 5847102.

Note: Patent information was last updated on 12 May 2006.
Search terms for Japan Tobacco patents: “promoter” in  abstract and “Japan Tobacco” in applicant.
Search terms for Danisco patents: “promoter” in  abstract and “Danisco” in applicant.
Search terms for Canadian Ministry of Agriculture patents: “promoter” in  abstract and “Canadian Ministry” in applicant.
Patent database: PatentLens in combination with INPADOC.