Quorum sensing signaling in bacteria

Abstract

The invention provides methods for identifying a modulator of quorum sensing signaling in bacteria, and for identifying a quorum sensing controlled gene in bacteria. In addition, the invention provides quorum sensing controlled genetic loci in Pseudomas aeruginosa. Novel indicator strains and vectors for engineering the strains for use in the method of the invention are also provided.

1 .- 45 . (canceled)  
     
     
         46 . An isolated nucleic acid molecule comprising a nucleotide sequence, said nucleotide sequence comprising: 
 a regulatory sequence derived from the genome of  Pseudomonas aeruginosa,  wherein said regulatory sequence regulates a quorum sensing controlled genetic locus of the  Pseudomonas aeruginosa  chromosome, and wherein said locus comprises a nucleotide sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36; and a reporter gene operatively linked to said regulatory sequence.    
     
     
         47 . An isolated nucleic acid molecule comprising a quorum sensing controlled genetic locus derived from the genome of  Pseudomonas aeruginosa,  wherein said locus comprises a nucleotide sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, operatively linked to a reporter gene.  
     
     
         48 . An isolated nucleic acid molecule comprising a polynucleotide having at least 80% identity to a quorum sensing controlled genetic locus derived from the genome of  Pseudomonas aeruginosa,  wherein said locus comprises a nucleotide sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, operatively linked to a reporter gene.  
     
     
         49 . An isolated nucleic acid molecule comprising a polynucleotide that hybridizes under stringent conditions to the complement of a nucleotide sequence comprising a quorum sensing controlled genetic locus derived from the genome of  Pseudomonas aeruginosa,  wherein said locus comprises a nucleotide sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ED NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, operatively linked to a reporter gene.  
     
     
         50 . The nucleic acid molecule of any one of claims  46 ,  47 ,  48  and  49 , wherein said reporter gene is contained in a transposable element.  
     
     
         51 . A vector comprising the isolated nucleic acid molecule of any one of claims  46 ,  47 ,  48  and  49 .  
     
     
         52 . A cell containing an isolated nucleic acid molecule of any one of claims  46 ,  47 ,  48  and  49 .  
     
     
         53 .- 74 . (canceled)  
     
     
         75 . The nucleic acid molecule of any one of claims  46 ,  47 ,  48  and  49 , wherein said reporter gene is lacZ, GFP, or a variant thereof.