Lyophilized bioluminescent bacterial reagent for the detection of toxicants
AssigneeE. I. Du Pont de Nemours and Company
InventorsTina Kangas Vandyk, Lorraine Winona Wagner
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Abstract
A reagent, useful in the detection of environmental insults comprising bacterial cells containing a stress promoter operably linked to a lux gene complex has been prepared by lyophilizing the cells in a specified medium. The reagent may be used immediately upon rehydration where a positive test for the presence of an environmental insult is given by an increase in light production from the cells.
Claims8
A method of detecting the presence of a sublethal environmental insult with a lyophilized biological reagent said reagent comprising a detector prokaryotic microorganism containing an expressible luxCDABE gene complex under the control of a stress inducible promoter sequence, the method comprising the steps of:
- (i) rehydrating the lyophilized biological reagent in a suitable amount of water wherein a baseline bioluminescence is produced;
- (ii) immediately contacting the rehydrated reagent with a sample suspected of containing an environment insult to form a reagent mixture;
- (iii) incubating the mixture for at least 20 minutes and at a temperature of up to 30.degree. C. and;
- (iv) detecting an increase in bioluminescence from the mixture.
A lyophilized biological reagent comprising bacteria transformed with an expressible luxCDABE gene complex under the control of a stress inducible promoter sequence.
A kit for detecting the presence of a sublethal environmental insult comprising the following in packaged combination:
- (i) an aliquoted lyophilized biological reagent comprising;
- (a) a detector prokaryotic microorganism cell containing a DNA fragment comprising a stress promoter gene operably linked to a luxCDABE gene complex;
- (b) a suitable buffer;
- and
- (c) a cryo protective reagent;
- (ii) a rehydrating reagent;
- and
- (iii) a suitable growth media.
Dependencies are inferred from the claim text.