| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 10824941 | Journal of Biochemical and Biophysical Methods | 2005 | 14 Pages |
Abstract
Escherichia coli DNA photolyase was expressed as C-terminal 6Ã histidine-fused protein. Purification of His-tagged E. coli DNA photolyase was developed using immobilized metal affinity chromatography with Chelating Sepharose Fast Flow. By one-step affinity chromatography, approximate 4.6 mg DNA photolyase was obtained from 400 ml E. coli culture. The purified His-tagged enzyme was combined with two chromophors, FADH and MTHF. Using the oligonucleotide containing cyclobutane pyrimidine dimer as substrate, both reversed-phase high-performance liquid chromatography and size-exclusion high-performance liquid chromatography were developed to measure the enzyme activity. The enzyme was found to be able to repair the cyclobutane pyrimidine dimer with the turnover rate of 2.4 dimers/photolyase molecule/min.
Keywords
Related Topics
Life Sciences
Biochemistry, Genetics and Molecular Biology
Biochemistry
Authors
Wanmeng Mu, Dongfang Zhang, Lei Xu, Zhaofeng Luo, Yuzhen Wang,