FIGURE 8.

Unique factor dependence of LSP relative to HSP1 when bubble templates are used. Proteins were assembled on the LSP1 or HSP1 bubble template (2.5 μm) by combining one or more of the following components: h-mtTFA (2.5 μm), h-mtTFB2 (2.5 μm), and h-mtRNAP (2.5 μm), as indicated, and ³²P-labeled dinucleotide primer (*pApA, 10 μm) in reaction buffer at 32 °C for 5 min. The reaction was initiated by addition of NTPs (500 μm each) in the presence or absence of heparin (10 μm) and then quenched at various times by addition of stop buffer. Products were resolved by denaturing PAGE on 25% gels and visualized by phosphorimaging. a, representative gel of an experiment using LSP bubble template. b, representative gel of an experiment using HSP1 bubble template.