Gene probes for the selective detection of microorganisms that reductively dechlorinate polychlorinated biphenyl compounds
OwnerKevin Sowers, Sonja Fagervoid +2inventors
InventorsKevin Sowers, Sonja Fagervoid, Joy Watts, Harold May
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Abstract
The present invention relates to an assay for identification of PCB dechlorinating organisms that are capable of biologically removing PCBs from contaminated materials. Specifically, the invention provides a set of primers for detecting PCB dechlorinating organisms in a sample. These individual primers of the primer set have a sequence of at least 12 nucleotides that is unique to 16S rDNA of PCB dechlorinating organisms.
Claims12
A set of primers for use in a detection assay for detecting PCB dechlorinating organisms in a sample, wherein the set of primers comprises SEQ ID NO:
- 1 and SEQ ID NO:
- 2 and any nucleotides sequences that are complementary to same, have more than 98% identity or that hybridize under high stringency conditions of 0.1×SSC, 0.1% SDS at 65° C.;
- and that hybridizes with 16S rRNA of a bacteria.
A method for identifying a PCB dechlorinating bacterial organism comprising:
- (i) extracting genomic DNA from a bacteria cell suspected of being able to dechlorinate PCB chlorinated compounds;
- (ii) probing the extracted genomic DNA with at least one probe having a sequence selected from the group consisting of:
- (a) SEQ ID NO:
- 1;
- (b) SEQ ID NO:
- 2;
- (c) a nucleotide sequence that is complementary to SEQ ID NO:
- 1 or SEQ ID NO:
- 2;
- (d) a nucleotide sequence having more than 98% identity with SEQ ID NO:
- 1 or SEQ ID NO:
- 2;
- (e) a nucleotide sequence that hybridizes under high stringency conditions of 0.1×SSC, 0.1% SDS at 65° C.;
- and that hybridizes with 16S rRNA of a bacteria,
- under suitable hybridization conditions, wherein the identification of a hybridizable nucleic acid fragment confirms the presence of a bacteria capable of dechlorinating PCB chlorinated compounds.
A method for the dechlorination of PCB chlorinated compounds comprising:
- contacting a PCB chlorinated compound with an isolated bacterial organism comprising a sequence consisting of SEQ ID NO:
- 4 under conditions suitable for PCB dechlorination to occur.
An isolated bacterial organism comprising a 16S ribosomal subunit nucleic acid sequence selected from the group consisting of:
- (a) a nucleic acid sequence that has more than 99% identity to a nucleic acid sequence of SEQ ID NO 4 and that hybridizes with 16S rRNA of a bacteria;
- and
- (b) a nucleic acid sequence fully complementary to a nucleic acid of (a);
- and
- wherein the isolated bioremediative microorganism anaerobically dechlorinates chlorinated biphenyls under conditions suitable for PCB dechlorination to occur.
A diagnostic nucleic acid gene fusion useful in Denaturing Gradient Gel Electrophoresis (DGGE) having the general structure:
- SS-GC, wherein:
- (i) SS is a signature sequence selected from the group consisting of SEQ ID NO:
- 1 and SEQ ID NO:
- 2;
- and
- (ii) GC is a GC clamp sequence having the sequence as set forth in SEQ ID NO:
- 5.
Dependencies are inferred from the claim text.