Isolation and analysis of CPD photolyase gene of spinach and fundamental study for the production pf transgenic plant resistant to UV

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We isolated a gene for cyclobutane pyrimidine dimer (CPD) photolyase in spinach and determined the function and the extent of the expression in male and female plants. cDNA of the gene was cloned by RTPCR with the use of niRNA as a tremplate and degenerate primers. The gene consists of nine exsons and amino acid sequence was quite similar to class II CPD photolyase. The gene obtined here restored the photoreactivating ability of mutant E. coil deficient in photorepair indicating that the gene was photolyase gene. Removal of CPD was confirmed by ELISA procedure. The gene was a single copy. The level of expression in root was lower than in leaves, stems and flowers in male and female spinach. UV generates reactive oxygen species (ROS). Specificity of ROS mutation was investigated in E. coil. Base change mutations were predominant among mutations.