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Patent applications filed by and patents owned by VITO 2

Technology overview

This series of patent documents by VITO (Vlaamse instelling voor technologisch onderzoek, Flemish Institute for Technological Research) describes novel SOS regulated promoters that can be used to express reporter genes that code for proteins that produce a signal that can be detected as light.  Promoters that were investigated for utility were wildtype and mutated recN promoters from E. coli, which were fused to plasmid pMOL877 containig a promoterless luxCDABE operon from Vibrio fischeri.  The specification compares their system to those of the SOS-Chromotest (genotoxicity test using E. coli containing a sfiA::lacZ fusion gene; Quillardet et al., Proc.Natl Acad. Sci. USA 1982, 79:5971-5975) and the Ames Test (mutagenicity test using S. typhimurium mutant strains; Maron and Ames, Mutat. Res. 1983, 113:173-215; Gee et al., Proc.Natl Acad. Sci. USA 1994, 91:11606-11610), and concludes that the novel promoter systems disclosed here is more sensitive and results can be obtained in a shorter amount of time.

The system disclosed in the patent documents in this section has been comercialized (later improved with introduction of a control strain; technology disclosed in another patent family filed by VITO – WO 1999/53092, see chapter 6: Patent applications filed and patents owned by VITO 3) as the VITOTOX test.  The related journal publication of this technology is van der Lelie et al. (1997).  The VITOTOX test, an SOS bioluminescence Salmonella typhimurium test to measure genotoxicity kinetics.  Mutat. Res. 389(2-3):279-90.

Details of patent documents

Patent or Publication no.

Title, Independent Claims and Summary

Assignee and licensing information
AU 724155

  • Earliest priority – 25 Apr 1996
  • Filed – 25 Apr 1996
  • Granted – 14 Sept 2000
  • Expected expiry – 25 Apr 2016
 Title – Recombinant nucleic acid sequences and methods for determining both genotoxicity and mutagenicity of a sample and the kinetics of the genetoxicity

Claim 1
A recombinant nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production.
Claim 20
A method for determining the presence of multiple genotoxic compounds in a sample, said method comprising the steps of- culturing a host microorganism, said host microorganism comprising a nucleic acid sequence comprising an SOS regulated promoter, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production,
– measuring the luminescence of the culture, said measuring occurring at various points in time, preferably continuously and determining the signal to noise ratio at said points in time, plotting the data, said data representing the kinetics of genotoxicity of said sample with multiple peaks being indicative of multiple genotoxicity compounds with different induction kinetics.
Claim 21
A method for determining the presence of a genotoxic compound in a sample, said method comprising the steps of- culturing a host microorganism, said host microorganism comprising a nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production,
– measuring the luminescence of the culture at multiple points in time, preferably continuously and
– determining whether the luminescence of the culture has changed, increased luminescence being indicative of the presence of a genotoxic compound.

The claims are generally drawn towards:

  • a recombinant nucleic acid sequence comprising an SOS regulated promoter operably linked to a reporter (claim 1)
  • a method for determining the presence of multiple genotoxic compounds in a sample (claim 20)
  • a method for determining the presence of a genotoxic compound in a sample (claim 21)

Definitions extracted from the description are:

  • SOS regulated promoter – there is no definition for this term. Candidates of suited promoters ‘from the recombinatorial repair promoters consist of the group RecF, RecJ, RecN, RecO, RecQ, ruv and uvrD promoters’
  • Induction ratio – Miller units (lexA71) / Miller units (lexA+), as stated in the provided reference in the specifications (Schnarr et al., Biochimie 1991, 73:423-431, which cites Peterson and Mount, J. Mol. Biol. 1987, 193:27-40). lexA71 is a mutation that results in the protein being deficient in DNA binding.
  • Reporter – there is no definition for this term. The description states that ‘such sequences are known in the state of the art’, and gives a suitable reporter gene as being the luciferase A and B genes, more preferably further comprising luciferase C, D, and E genes, the sequences of which the description refers to that of WO 1992/15687 (see chapter 5 – Patent applications filed and patents owned by VITO).

Comments:

Although the description states that the SOS regulated promoter in the subject invention is ‘directed at a recombinant nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40’, independent claim 20 does not state the limitaion of the SOS regulated promoter as such (in contrast to claims 1 and 21, see above), therefore may encompass any SOS regulated promoter.

 VITO

BOERETANG 200
B-2400
MOL BELGIUM

Ph + 32 14 33 55 11
Fax + 32 14 33 55 99

vito@vito.be
CA 2252699

  • Earliest priority – 25 Apr 1996
  • Filed – 25 Apr 1996
  • Application pending
  • Expected expiry – not applicable
 Title – Recombinant nucleic acid sequences and methods for determining both genotoxicity and mutagenicity of a sample and the kinetics of the genetoxicity

Claim 1
A recombinant nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production.
Claim 20
A method for determining the presence of multiple genotoxic compounds in a sample, said method comprising the steps of- culturing a host microorganism, said host microorganism
comprising a nucleic acid sequence comprising an SOS regulated promoter, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production,
– measuring the luminescence of the culture, said
measuring occurring at various points in time, preferably continuously and determining the signal to noise ratio at said points in time, plotting the data, said data representing the kinetics of genotoxicity of said sample with multiple peaks being indicative of multiple genotoxicity compounds with different induction kinetics.
Claim 21
A method for determining the presence of a genotoxic compound in a sample, said method comprising the steps of- culturing a host microorganism, said host microorganism comprising a nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 20, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production,
– measuring the luminescence of the culture at multiple points in time, preferably continuously and
– determining whether the luminescence of the culture has changed, increased luminescence being indicative of the presence of a genotoxic compound.

The claims are generally drawn towards:

  • a recombinant nucleic acid sequence comprising an SOS regulated promoter operably linked to a reporter (claim 1)
  • a method for determining the presence of multiple genotoxic compounds in a sample (claim 20)
  • a method for determining the presence of a genotoxic compound in a sample (claim 21)

Definitions extracted from the description are provided in AU 724155 (see above).

Comments:

Since this is a published application and not a granted patent, currently there are no enforceable rights.
US 6521751

  • Earliest priority – 25 Apr 1996
  • Filed – 25 Apr 1996
  • Granted – 18 Feb 2003
  • Expected expiry – 25 Apr 2016
 Title – Recombinant nucleic acid sequences and methods for determining both genotoxicity and mutagenicity of a sample and the kinetics of the genetoxicity

Claim 1
A recombinant nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production, wherein the promoter is Rec N (seq. id. no. 3).
Claim 2
A recombinant nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production, wherein the promoter is a mutated RecN promoter with the following nucleic acid sequence id. no. 9.
Claim 3
A recombinant nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production, wherein the promoter is a mutated RecN promoter with the following nucleic acid sequence id. no. 10.

The claims are generally drawn towards:

  • a recombinant nucleic acid sequence comprising RecN operably linked to a reporter (claim 1, 2, 3)

Definitions extracted from the description are provided in AU 724155 (see above). The SOS regulated promoter is limited to those described as SEQ ID NOs:3, 9 and 10.

  • SEQ ID NO: 3 – wildtype recN promoter DNA sequence
  • SEQ ID NO: 9 – recN promoter DNA sequence lacking the LexA2 site (recN1-3)
  • SEQ ID NO: 10 – recN promoter DNA sequence with promoter up mutation (recN2-4)
  • Promoter up mutation – the mutation is such that the promoter sequence more closely resembles that of the consensus sequence for the RNA polymerase binding site.

Comments:

US 6521751 has the narrowest set of independent claims among its patent family, as the nucleic acid sequence of the SOS regulated promoter is limited to three sequences (SEQ ID NOs: 3, 9, and 10) coding for a wildtype (SEQ ID NO:3) and mutated (SEQ ID NOs:9 and 10) RecN.
WO 1997/41251

  • Earliest priority – 25 Apr 1996
  • Filed – 25 Apr 1996
  • Published – 6 Nov 1997
  • Expected expiry – not applicable
 Title – Recombinant nucleic acid sequences and methods for determining both genotoxicity and mutagenicity of a sample and the kinetics of the genetoxicity

Claim 1
A recombinant nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 40, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production.
Claim 20
A method for determining the presence of multiple genotoxic compounds in a sample, said method comprising the steps of- culturing a host microorganism, said host microorganism comprising a nucleic acid sequence comprising an SOS regulated promoter, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production,
– measuring the luminescence of the culture, said measuring occurring at various points in time, preferably continuously and determining the signal to noise ratio at said points in time, plotting the data, said data representing the kinetics of genotoxicity of said sample with multiple peaks being indicative of multiple genotoxicity compounds with different induction kinetics.
Claim 21
A method for determining the presence of a genotoxic compound in a sample, said method comprising the steps of- culturing a host microorganism, said host microorganism comprising a nucleic acid sequence comprising an SOS regulated promoter with an induction ratio higher than 20, said promoter being operatively linked to a reporter encoding nucleic acid sequence encoding a reporter resulting in a signal that can be assayed as light production,
– measuring the luminescence of the culture at multiple points in time, preferably continuously and
– determining whether the luminescence of the culture has changed, increased luminescence being indicative of the presence of a genotoxic compound.

The claims are generally drawn towards:

  • a recombinant nucleic acid sequence comprising an SOS regulated promoter operably linked to a reporter (claim 1)
  • a method for determining the presence of multiple genotoxic compounds in a sample (claim 20)
  • a method for determining the presence of a genotoxic compound in a sample (claim 21)

Definitions extracted from the description are provided in AU 724155 (see above).

Comments:

Since this is a published application and not a granted patent, currently there are no enforceable rights.
Remarks
  1. National phase entry of WO 1997/41251 in Japan (JP 2001/507922) was rejected on 4 Apr 2006.  No further correspondence has been reported as of 18 may 2006.
  2. National phase entry of WO 1997/41251 in Europe (EP 907748) has deemed to be withdrawn on 24 Aug 2005.
  3. Other national phase entry of WO 1997/41251 include Brazil (BR 9612617).

Search strategy

Search details
Date of search 17/05/2006
Database searched Patent Lens
Type of search Simple, stemming on
Collections searched AU-B, US-A, US-B, EP-B, WO
Search terms VITO in applicant
Results 3
Comments Of the 3 results identified using these search terms, 3 results were identified as being of particular interest based on their abstracts and a review of their claims.