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Cloning, sequencing, expression and characterization of DNA photolyase from Salmonella typhimurium.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Li, Yuling Sancar, Aziz |
| Copyright Year | 1991 |
| Abstract | We have cloned the phr gene that encodes DNA photolyase from Salmonella typhimurium by in vivo complementation of Escherichia coli phr gene defect. The S.typhimurium phr gene is 1419 base pairs long and the deduced amino acid sequence has 80% identity with that of E. coli photolyase. We expressed the S.typhimurium phr gene in E.coli by ligating the E.coli trc promoter 5' to the gene, and purified the enzyme to near homogeneity. The apparent molecular weight of S.typhimurium photolyase is 54,000 dalton as determined by SDS-polyacrylamide gel electrophoresis, which is consistent with the calculated molecular weight of 53,932 dalton from the deduced phr gene product. S.typhimurium photolyase is purple-blue in color with near UV-visible absorption peaks at 384, 480, 580, and 625 nm and a fluorescence peak at 470 nm. From the characteristic absorption and fluorescence spectra and reconstitution experiments, S.typhimurium photolyase appears to contain flavin and methenyltetrahydrofolate as chromophore-cofactors as do the E.coli and yeast photolyases. Thus, S.typhimurium protein is the third folate class photolyase to be cloned and characterized to date. The binding constant of S.typhimurium photolyase to thymine dimer in DNA is kD = 1.6 x 10(-9) M, and the quantum yield of photorepair at 384 nm is 0.5. |
| Starting Page | 760 |
| Ending Page | 762 |
| Page Count | 3 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://oup.silverchair-cdn.com/oup/backfile/Content_public/Journal/nar/19/18/10.1093/nar/19.18.4885/2/19-18-4885.pdf?Expires=1493097450&Key-Pair-Id=APKAIUCZBIA4LVPAVW3Q&Signature=B4e1-Qwb6~FtKGGCGyHlswSLKCZrLIBzC1F9S59ERl3dBCgEQhL~yaeX7eHVJZnYdW2HrrpGz3e1Ix0ZaF1v7kJIZUKlH-l4~1ipnnKS2m7zx3Uuj9UtTU92hsm6Wen-lZOohXWjcfsx0wL8ZdtKnKmAcsPsiORG385nOxzclSGwyXoGIk7SFEknfeFMRVrcouCqlKmzrNtirpcizGF1jPsJUKR28g0-bv-e9tABCx7pmMmjbxX~sjZVgeaW2d2gRCeaJL3PjAH8uRqbru2XweyoBNYEIDN6d-mqNIMZLtutZEx26e4mhGGqe6OgTYF3kRIV~OPKYks-7UXdP6895Q__ |
| PubMed reference number | 1840665v1 |
| Volume Number | 19 |
| Issue Number | 18 |
| Journal | Nucleic acids research |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | 4,6-dinitro-o-cresol Amino Acid Sequence Amino Acids Base Pairing Biopolymer Sequencing Clone Cells DNA Photolyase Eighty Fluorescence Folic Acid Gel Electrophoresis (lab technique) Genome Encoded Entity Molecular Weight Polyacrylamide Gel Electrophoresis Salmonella typhimurium H Ab:Titr:Pt:Ser:Qn Salmonella typhimurium H Ab:Titr:Pt:Ser:Qn:Aggl Thymine UV-VIS Spectrum chemical cofactor cyclobutane pyrimidine dimer photolyase, Cucumis sativus dalton polyacrylamide gels |
| Content Type | Text |
| Resource Type | Article |
