Highly sensitive method for detecting environmental insults
AssigneeE. I. Du Pont de Nemours and Company
InventorsRobert Alan LaRossa, William Robert Majarian, Tina Kangas Van Dyk
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Abstract
Subtle changes in environmental stress can now be detected and measured at sublethal levels as a generic response to environmental stress. The present invention provides a method to detect changes in the environmental stress level. The stress change is indicated as a change in the luminescence output of a genetically engineered microorganism. In the present invention, the luminescence gene complex is under the control of a stress inducible promoter.
Claims13
A method of detecting the presence of an environmental insult comprising:
- (a) exposing a transformed detector E. coli to an environmental insult, the transformed detector E. coli being genetically engineered to contain an expressible heterologous luxCDABE gene complex under the control of a stress-inducible promoter sequence wherein the promoter sequence is responsive to a regulatory circuit;
- and
- (b) measuring an increase in luminescence of the transformed detector E. coli, the increase indicating the presence of an environmental insult.
A method of detecting stress in a population of transformed E. coli comprising:
- (a) exposing a population of transformed detector E. coli to an environmental insult, the transformed detector E. coli being genetically engineered to contain an expressible heterologous luxCDABE gene complex trader the control of a stress-inducible promoter sequence wherein the promoter sequence is responsive to a global regulatory circuit;
- and
- b) measuring an increase in luminescence of the transformed detector E. coli, the increase indicating stress.
A transformed bioluminescent E coli capable of an increase in bioluminescence upon exposure to a sublethal level of environmental insult, the transformed bioluminescent E. coli comprising:
- (a) a stress inducible promoter sequence wherein the promoter sequence is responsive to a regulatory circuit;
- and
- (b) an expressible heterologous luxCDABE gone complex under the control the stress inducible promoter sequence.
A method of detecting the presence of a environmental insult comprising:
- (a) exposing a transformed detector E. coli to a sublethal environmental insult, the transformed detector E. coli being genetically engineered to contain an expressible heterologous luxCDABE gone complex under the control of a stress-inducible promoter sequence wherein the promoter sequence is responsive to a regulatory circuit;
- and
- (b) measuring an increase in luminescence of the transformed detector E. coli, the increase indicating the presence of an environmental insult.
E. coli selected from the group consisting of:
- (i) TV1076 having ATCC Number 69314 comprising a tolC.sup.- mutation and an expressible heterologous lux gene complex under the control of a grpE stress inducible promoter sequence;
- (ii) WM1302 having ATCC Number 69316 comprising a tolC.sup.- mutation and an expressible heterologous lux gene complex under the control of a dnaK stress inducible promoter sequence;
- (iii) TV1060 having ATCC Number 69142 comprising an expressible heterologous lux gene complex under control of a grpE stress inducible promoter sequence;
- (iv) TV1061 having ATCC Number 69315 comprising an expressible heterologous lux gene complex under control of a grpE stress inducible promoter sequence;
- (v) WM1021 having ATCC Number 69141 comprising an expressible heterologous lux gene complex under control of a dnaK stress inducible promoter sequence;
- (vi) WM1026 having ATCC Number 69143 comprising an expressible heterologous lux gene complex under control of a dnaK stress inducible promoter sequence;
- and
- (vii) WM1202 having ATCC Number 69313 comprising an expressible heterologous lux gene complex under control of a dnaK stress inducible promoter sequence.
A nucleic acid molecule, comprising:
- (a) a stress inducible promoter sequence wherein said promoter sequence is responsive to a regulatory circuit;
- and
- (b) an expressible bacterial luxCDABE gene complex under control of said promoter sequence.
Dependencies are inferred from the claim text.