Compositions and methods for regulating bacterial pathogenesis
View Patent ↗The production of a purified extracellular bacterial signal called autoinducer-2 is regulated by changes in environmental conditions associated with a shift from a free-living existence to a colonizing or pathogenic existence in a host organism. Autoinducer-2 stimulates LuxQ luminescence genes, and is believed also to stimulate a variety of pathogenesis related genes in the bacterial species that produce it. A new class of bacterial genes is involved in the biosynthesis of autoinducer-2.
1. A method for detecting an autoinducer, selected from the group consisting of 4,5-dihidroxy-2,3-pentanedione (AI-2) and an analog thereof capable of interacting with Vibrio haveyi LuxP or LuxQ protein and thereby inducing Vibrio harveyi luminescence, in a sample comprising:
a) contacting the sample with a bacterial cell capable of producing a detectable amount of light in response to the autoinducer, wherein the bacterial cell comprises a first alteration in a sensor gene locus that inhibits detection of a first autoindecuer and a second alteration in a biosynthetic gene locus that inhibits production of 4,5-dihidroxy-2,3-pentanedione (AI-2) or an analog thereof capable of interacting with Vibrio harveyi LuxP or LuxQ protein and thereby inducing Vibrio harveyi luminescence; and
b) detecting light produced by the bacterial cell, thereby detecting an exogenous autoinducer in the sample.
2. The method of claim 1 , wherein the first alteration inhibits detection of (N-(3-hydroxybutanoyl)-L-homoserine lactone) (AI-1).
3. The method of claim 2 , wherein the first bacterial cell is luxN − .
4. The method of claim 3 , wherein the bacterial cell is luxS − .
5. The method of claim 4 , wherein the bacterial cell is V. harveyi strain MM32 (ATCC access No. BAA-1121.