Nucleotide Excision Repair in Nuclear Extracts from Xenopus Oocytes
Limited nucleotide excision repair (NER) requires at least {approx}40 proteins in extracts from purified proteins, although perhaps hundreds of proteins may influence DNA repair in cells. For efficient DNA repair in extracts, it is important to utilize a system containing large quantities of active DNA repair proteins uncontaminated with nonspecific nucleases. Unlike extracts from mammalian cells that repair {approx}2% of the input DNA, both injected Xenopus oocytes and oocyte nuclear extracts can repair {approx}100% of the input damaged DNA by NER with little or no synthesis on undamaged control substrate. Repair activity in extracts can be inactivated with antibodies and/or inhibitors, and then repair can be restored by addition of exogenous proteins. A further advantage of the Xenopus system is that results obtained from injection experiments in living cells can be compared to results obtained in nuclear extracts.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 896082
- Report Number(s):
- PNWD-SA-5367; TRN: US200703%%497
- Resource Relation:
- Related Information: Methods in Molecular Biology: DNA Repair Protocols; Eukaryotic Systems, 113:347-355
- Country of Publication:
- United States
- Language:
- English
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