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Top1-Clamp is catalytically active in vivo. Top1, Top1-Clamp, and Top1G365C activity were assayed in top1Δ, top2ts yeast expressing bacterial topA as described (12). The distribution of 2-μm DNA topoisomers was determined by Southern blotting of 2D gels. Along the topoisomer arc, positively (+) and negatively (–) supercoiled DNAs are shown. The pGAL vector was used as a negative control.
