Patent applications filed by The University of Tennessee Research Corporation

Technology overview

The patent family described in this section was filed by a research team lead by Dr John Sanseverino and Dr Gary S. Sayler at the Center for Environmental Biotechnology at the University of Tennessee, Knoxville.  They have identified a method to detect the presence of divalent mercury by transforming E. coli and P. fluorescens with the operator/promoter (o/p) region of merR (merRo/p) fused to luxCDABE using a transposable element to integrate the fusion gene into the bacterial genome.  They have also investigated the use of a support matrix and an encapsulating material for the transgenic bacterium to provide a user-friendly detection kit for mercury, and disclosed the technology.  A related scientific presentation was held by the research group in 2001 – Carrasquillo CI, Tomaszewski A, Sanseverino J, Sayler GS (2001).  Development of a handheld device for the detection of divalent mercury in water using a genetically engineered reporter microorganism.  Society for the Advancement of Chicanos and Native Americans in Science. 27-30 Sept, Phoenix, Arizona.

Details of patent documents

Patent or Publication no. Title, Independent Claims and Summary Assignee and licensing information
CA 2419481

  • Earliest priority – 14 Aug 2000
  • Filed – 10 Aug 2001
  • Application pending
  • Expected expiry – not applicable
Title – Bioluminescent methods for direct visual detection of environmental compounds

Claim 1
A device for detecting a selected analyte, comprising:

  • a stably transformed bacterium containing a promoterless lux gene cassette having a regulatory element for a selected analyte inserted in front of the lux gene cassette;
  • a support matrix onto which the bacterium is attached; and
  • an encapsulating material to contain said bacterium attached to the support matrix wherein the encapsulated bacterium emits visibly detectable light in the presence of the selected analyte.
Claim 13
A genetically modified bacterium responsive to divalent mercury, said bacterium containing a merRo/p-lux gene stably integrated into the bacterial chromosome wherein said bacterium produces a bioluminescent protein in the presence of divalent mercury.
Claim 25
A method for detecting mercury comprising

  • contacting a sample suspected of containing mercury II ion with a bioreporter bacterium genetically modified to contain a merRo/pA-lux gene; and
  • detecting the presence of the mercury ion when a visibly detectable luminescence is produced.

The claims are generally drawn towards:

  • a device for detecting a selected analyte comprising a bacterium containing a promoterless lux gene cassette having a regulatory element for a selected analyte inserted in front of the lux gene cassette (claim 1)
  • a genetically modified bacterium responsive to divalent mercury (claim 13)
  • a method for detecting mercury comprising contacting a sample with a bioreporter bacterium genetically modified to contain a merRo/p-lux gene (claim 25)

Definitions extracted from the specification are provided in WO 2002/14551.

Comments:

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

University of Tennessee Research Corporation

1534 WHITE AVENUE
SUITE 403
KNOXVILLE, TENNESSEE 37996
Ph: +1-865-974-1882

Email: vhunley@tennessee.edu

US 2003/108980

  • Earliest priority – 14 Aug 2000
  • Filed – 8 Aug 2001
  • Application abandoned – 15 Mar 2006
  • Expected expiry – not applicable
Title – Bioluminescent methods for direct visual detection of environmental compounds

Claim 1
A device for detecting a selected analyte, comprising:

a stably transformed bacterium containing a promoterless lux gene cassette having a regulatory element for a selected analyte inserted in front of the lux gene cassette;
a support matrix onto which the bacterium is attached; and
an encapsulating material to contain said bacterium attached to the support matrix wherein the encapsulated bacterium emits visibly detectable light in the presence of the selected analyte.

Claim 13
A genetically modified bacterium responsive to divalent mercury, said bacterium containing a merRo/p-lux gene stably integrated into the bacterial chromosome wherein said bacterium produces a bioluminescent protein in the presence of divalent mercury.
Claim 25
A method for detecting mercury comprising

  • contacting a sample suspected of containing mercury II ion with a bioreporter bacterium genetically modified to contain a merRo/pA-lux gene; and detecting the presence of the mercury ion when a visibly detectable luminescence is produced.

The claims are generally drawn towards:

  • a device for detecting a selected analyte comprising a bacterium containing a promoterless lux gene cassette having a regulatory element for a selected analyte inserted in front of the lux gene cassette (claim 1)
  • a genetically modified bacterium responsive to divalent mercury (claim 13)
  • a method for detecting mercury comprising contacting a sample with a bioreporter bacterium genetically modified to contain a merRo/p-lux gene (claim 25)

Definitions extracted from the specification are provided in WO 2002/14551.

Comments:

This application has been abandoned due to failure of the applicant to respond to an office action.

Because it is a published application and not a granted patent, there are no enforceable rights.

WO 2002/14551

  • Earliest priority – 14 Aug 2000
  • Filed – 10 Aug 2001
  • Published – 21 Feb 2002
  • Amended claims – 26 Jun 2003
Title – Bioluminescent methods for direct visual detection of environmental compounds

Claim 1
A portable system for detecting a selected analyte, comprising:

  • a stably transformed bacterium containing a promoterless lux gene cassette having a regulatory element for a selected analyte inserted in front of the lux gene cassette;
  • support matrix onto which the bacterium is attached ;
  • an encapsulating material to contain said bacterium attached to the support matrix, wherein the encapsulated bacterium emits visibly detectable light in the presence of the selected analyte, and
  • a portable detection device.
Claim 13
A genetically modified bacterium responsive to divalent mercury, said bacterium being encapsulated and containing a gene stably integrated into bacterial chromosome, wherein said bacterium produces a bioluminescent protein in the presence of divalent mercury.
Claim 22
A mobile method for detecting mercury in water samples comprising

  • providing a plurality of stably transformed bioreporter bacterium genetically modified to contain a merRo/p-lux gene, said bacterium attached to a support matrix and disposed within protective packaging for preserving hydration of said bacterium;
  • removing said protective packaging;
  • contacting a water comprising sample suspected of containing mercury II ion with said detecting the presence of the mercury ion when a visibly detectable luminescence is produced, said detecting using a portable detection device.

Note: The three independent claims extracted above are those of the amended claims.

The claims are generally drawn towards:

  • a portable system for detecting a selected analyte comprising a bacterium containing a promoterless lux gene cassette having a regulatory element for a selected analyte inserted in front of the lux gene cassette (claim 1)
  • a genetically modified bacterium responsive to divalent mercury (claim 13)
  • a mobile method for detecting mercury in water samples comprising providing a bioreporter bacterium genetically modified to contain a merRo/p-lux gene (claim 22)

Definitions extracted from the specification are:

  • Bioreporter – intact, living microbial cells that have been genetically engineered to produce a measurable signal in response to a specific chemical or physical agent in their environment.
  • lux gene cassette – this term is not defined in the specification. The preferred embodiment is provided as follows: ‘…the cassette may be merRo/p-lux where the lux gene comprises CDABE.’
  • Analyte – this term is not defined in the specification. Dependent claims 4 and 5 recite the following analytes: naphthalene, toluene, ethylbenzene, 2,4-dichlorophenoxyacetic acid, 13-phenylethylamine, phenol, biphenyl, and mercury.
  • Bacterium – the following statement indicates that there is no limit on the bacterial species that can be used for this invention: ‘While the invention has been illustrated with E. coli and P. fluorescens, other bacteria can be used.’
  • Support matrix – this term is not defined in the specification. Dependent claim 17 recite the following matrices: cellulose, glass, colloidal noble metal, plastic, laminin or resin.

Comments:

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

Examples in the specification have described the enablement of the technology by:

  • Creating a merRo/p::lux bioreporter – merRo/p (Genbank Accession AF071413; nucleotides 19133-19638) was fused to luxCDABE on pFSP3, the mer-lux reporter transposon was excised to create a transposome, which was then electroporated into E. coli EC 100 competent cells. Three transformants gave bioluminescence in response to Hg++, one of which had twice luminescence intensity compared to the other two strains.
  • Immobilising the bioreporter – toxity testing of latex material and bioluminescence of immobilised bioreporters were designed with P. fluorescens 5RL (naphthalene detection). A simulated bioluminescene test with bioreporters immobilised in Noble agar onto  a layer of silicone plastic was conducted with several bioreporter strains, including that of E. coli EC 100 containing merRo/p::lux.
Remarks
  1. WO 2002/14551 has entered national phase in the United States, which has not yet been published.
  2. National phase entry of WO 2002/14451 in Australia (AU 2001/86427) lapsed on 10 Jan 2004.
  3. National phase entry of WO 2002/14451 in Europe (EP 1315836) is pending.

Search strategy

Search details
Date of search 14/09/2006
Database searched Patent Lens
Type of search Simple, stemming on
Collections searched AU-B, US-A, US-B, EP-B, WO
Search terms bioreporter and (Sayler in inventor)
Results 27
Comments Of the 27 results identified using these search terms, 4 results were identified as being of particular interest based on their abstracts and a review of their claims.