Production of cis-syn thymine-thymine cyclobutane dimer oligonucleotide in the presence of acetone photosensitizer.
cis-syn Cyclobutane pyrimidine dimer (CPD) oligonucleotide was produced by UV irradiation in the presence of acetone photosensitizer. Acetone could enhance the productivity but evidently induced the photocleavage of oligonucleotide under a long time irradiation. A statistical approach of orthogonal design was applied to optimize the preparation condition for the production of the modified oligonucleotide. Optimal conditions for maximal cis-syn CPD oligonucleotide productivity were determined based on three factors: acetone concentration, initial oligonucleotide concentration, and irradiation time at several different levels. The optimal modified oligonucleotide that this optimization could produce was 32.7%. Through analysis of 20% polyacrylamide gel electrophoresis, it was found that modified oligonucleotide migrated slightly more slowly than the parent oligonucleotide. The photoreactivation of cis-syn thymine-thymine dimer oligonucleotide displayed the selectivity of the substrate specificity of DNA photolyase with high-performance liquid chromatography (HPLC) analysis.
School of Life Sciences, University of Science and Technology of China, Hefei.
Wanmeng
W
Mu
Qingkai
Q
Han
Zhaofeng
Z
Luo
Yuzhen
Y
Wang
D000096:Acetone / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D019281:Dimerization; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry; D010777:Photochemistry / Q000379:methods*; D017319:Photosensitizing Agents / Q000737:chemistry*; D011740:Pyrimidine Dimers / Q000138:chemical synthesis* / Q000737:chemistry; D012107:Research Design; D014466:Ultraviolet Rays
D009841:Oligonucleotides; D017319:Photosensitizing Agents; D011740:Pyrimidine Dimers; D000096:Acetone