In Vitro Excision Repair Assay in Schizosaccharomyces pombe

The measurement of DNA excision repair activity in vitro, originally developed by Wood and co-workers (1 ), utilizes transcriptionally active protein extracts obtained from mammalian cells by the method of Manley et al. (2 ) (seeChapter 29 ). Nucleotide excision repair (NER), which requires more than 20 proteins in vitro, can process a large variety of lesions according to the following mechanism:

1. Recognition of the DNA lesion.
2. Incision on both sides of the lesion.
3. Excision of the damaged oligonucleotide.
4. DNA polymerization and ligation.

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