Fig. 2. HO-inducible SSA assay.
(A) Overview of the HO-inducible SSA assay. HOcs denotes HO cleavage site. A downward arrow indicates the cleavage by HO endonuclease. Horizontal arrows indicate the position and direction of oligonucleotides used for PCR amplification. The sequences are as follows: Forward: 5'-GAC TCA TGG CAA GAT TTC TC-3'; Reverse: 5'-AAG TCA AGT GAG GTA TGT AG-3'. In this assay, DSB can be repaired by NHEJ or SSA. NHEJ restores HOcs leaving URA3 intact, whereas SSA removes URA3, generating a chimeric ASN gene. (B) SSA frequencies in the presence (grey squares) and absence (open square) of galactose induction. SSA frequency was calculated by dividing the number of colonies growing on SD plates containing 5-FOA by the number of colonies growing on YPD plates. The results are the average (± SD) of 5 independent experiments. Asterisks indicate statistical significance (*P < 0.001).
© 2020 Korean J. Microbiol.
