Patent ID: 6921641
Filing Date: 2005-07-26
Classification: C12N

Abstract:
1. A method for detecting DNA damage, the method comprising: combining a first polypeptide, a second polypeptide and a double stranded DNA to form a mixture; wherein the first polypeptide is encoded by a first polynucleotide, wherein the complement of the first polynucleotide hybridizes to SEQ ID NO:1 under standard hybridization conditions, and wherein the first polypeptide forms a complex at about 50° C. to about 80° C., the complex comprising the first polypeptide, a UvrB polypeptide comprising SEQ ID NO:4, and a BPDE-DNA substrate; wherein the second polypeptide is encoded by a second polynucleotide wherein the complement of the second polynucleotide hybridizes to SEQ ID NO:3 under standard hybridization conditions, and wherein the second polypeptide forms a complex at about 50° C. to about 80° C., the complex comprising the second polypeptide, a UvrA polypeptide comprising SEQ ID NO:2, and a BPDE-DNA substrate; incubating the mixture such that a complex forms comprising the first polypeptide, the second polypeptide, and the double stranded DNA; detecting the complex, wherein the presence of a complex indicates the presence of DNA damage; wherein standard hybridization conditions are 6×SSC, 5×Denhardt, 0.5% sodium dodecyl sulfate (SDS), and 100 μg/ml fragmented and denatured salmon sperm DNA hybridized overnight at 65° C. and washed in 2×SSC, 0.1% SDS at least one time at room temperature for about 10 minutes followed by at least one wash at 65° C. for about 15 minutes followed by at least one wash in 0.2×SSC, 0.1% SDS at room temperature for at least 3-5 minutes.