Systematic Quantification of GFP-tagged Protein Foci in Schizosaccharomyces pombe Nuclei.
Bio Protoc
; 8(24): e3117, 2018 Dec 20.
Article
em En
| MEDLINE
| ID: mdl-34532559
ABSTRACT
DNA damage repair proteins form foci in response to DNA damaging agents. The efficiency and integrity of the DNA repair pathway of a particular eukaryotic (mutant) strain is usually determined by the number of foci formed compared with their wild-type counterpart. Conventionally, focus number is determined visually, and this low accuracy may obscure the identification of a weaker phenotype, particularly when the output is low. Here, using the homologous recombination protein Rhp54 as an example, we present a protocol that can increase the consistency of foci identification among samples and can significantly improve the efficiency of foci quantification for large sample sizes. A similar method can be applied to other foci-forming proteins.
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MEDLINE
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En
Ano de publicação:
2018
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Article