In Vitro Excision Repair Assay in Schizosaccharomyces pombe
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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 ) (see Chapter 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.
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| 2. |
Incision on both sides of the lesion.
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| 3. |
Excision of the damaged oligonucleotide.
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| 4. |
DNA polymerization and ligation.
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