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Competition between interchromosomal and intrachromosomal donors.

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posted on 11.07.2017, 17:37 by Ruoxi W. Wang, Cheng-Sheng Lee, James E. Haber

(A) Scheme for DSB repair in diploid strains. The DSB could be repaired by gene conversion using an ectopic intrachromosomal LEU2 sequence, an allelic leu2-KpnI sequence, or the homologous sequence outside of the leu2-KpnI. Each outcome is revealed by KpnI-digestion of the indicated PCR fragments, yielding respectively 3, 2 and 1 fragments, as illustrated in (C). (B) Usage of ectopic and allelic donors assessed from 31 colonies of individual recombinants in YWW210 (58% intrachromosomal donor usage assessed in pooled cells from S5 Fig and 85% viability in haploid strain from Fig 1). (C) Three types of possible outcomes for individual repair events revealed by KpnI-digestion: (1) repair from intrachromosomal LEU2; (2) allelic repair without URA3 co-conversion; (3) allelic repair with URA3 co-conversion.