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. 2005 Aug 15;19(16):1905–1919. doi: 10.1101/gad.337205

Figure 1.

Figure 1.

Using the Fob1–RFB system to pause specific DNA replication forks on chromosome 3. (A, panel i) Map of two consecutive repeats of the rDNA on chromosome 12 of budding yeast. The positions of the replication fork barrier (RFB), origin of DNA replication (ARS), 35S rRNA (35S), and 5S rRNA (5S) genes are indicated. (Panel ii) Insertion of RFB sequences on chromosome 3 to block the forks from ARS305 and ARS306. The distances indicated are measured from the left end of chromosome 3; “B” and “S” mark the positions of BclI and SalI restriction enzyme sites used for the two-dimensional DNA gels. See Materials and Methods and Supplementary Figure 1 for more information. (Panel iii) Using two-dimensional DNA gels to study DNA replication intermediates; see text for details. The two spots below the Y-arc in the left panel correspond to other sites in the genome that are recognized weakly by the chosen probe. (B) DNA replication forks do not pause at the RFBs on chromosome 3 in the absence of Fob1. (C, panel i) In cells expressing Fob1, the forks from ARS305 and ARS306 pause for an extended period at the two RFBs on chromosome 3. (Panel ii) The histograms show a quantification of paused forks at the indicated times (calculated as described in Materials and Methods).