Identification of genomic targets of DNA-binding proteins using Ty5. (A) Sir4 fused to a DNA-binding protein causes Ty5 to integrate into the genome near the binding sites for that transcription factor (TF). (B) After Ty5 transposition (➀), genomic DNA is cleaved with a restriction enzyme that cuts near the end of Ty5 (➁) and is ligated in dilute solution to favor recircularization of the fragments (➂). This is followed by amplification of the circular DNA that contains the end of the transposon and flanking genomic DNA by an “inverse PCR” (➃), and the identity of the flanking genomic DNA is determined by DNA sequencing or hybridization to a DNA microarray (➄).
