Figure 5.

Capture of DNA fragments from higher eukaryotes at fission yeast DSBs. (A) Frequency of ura4⁺ molecules with inserted DNA after transformation of exponentially growing wild-type (PN559), rhp51Δ (PN2490), rad50Δ (KT120), lig4Δ (AD459), and pku70Δ (PN3773) cells with 2 μg ura4⁺ DNA and 50 μg sheared salmon sperm DNA. The number of ura4⁺ molecules analyzed is shown on the bars. Three to six independent transformations were performed for each strain. (B) Frequency of fission yeast mtDNA in ura4⁺ inserts recovered from cells transformed with 2 μg ura4⁺ DNA only (−) and either 50 μg salmon sperm DNA or 50 μg human genomic DNA. (C) Inserts recovered from wild-type and rhp51Δ cells were classified as follows: fission yeast mtDNA, salmon microsatellite DNA, salmon repetitive DNA including ribosomal DNA, and other salmon DNA sequences. (D) Characteristics of human genomic DNA inserts recovered in the assay. (E) Insert of clone 4 showing the presence of (GT)₁₇ repeats at the 3′ junction. WT, wild type.