FIG. 6.

Cloned Tf2-neo transpositions. (A) Sequences of two transpositions with flanking genomic DNA. Tf2-neo candidate transpositions were cloned into pBSIISK−, and DNA flanking the event was sequenced, by using both the Tf2 and pBSII primers. WT22 shows the sequence obtained directly flanking either end of Tf2-neo, including TSDs (underlined). For WT3, the sequence flanking the 5′ LTR, including the putative TSD, was obtained; however, the sequence immediately flanking the 3′ LTR actually represents the predicted sequence at the site of insertion, based on identity of the 5′ flanking sequence to an S. pombe sequence in the Sanger Centre database. Thus, the indicated TSD and 3′ flanking sequence is predicted. (B) YHL912 sequences at Tf2-neo insertion sites. By using the S. pombe sequence database, the target site for each transposition event was found to lack Tf2 homology, having neither a Tf2 LTR nor a full-length Tf2; this was confirmed by PCR analysis of the genomic site of insertion (data not shown). Thus, WT22 and WT3 represent true transposition events.