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Figure 3 | Mobile DNA

Figure 3

From: Protein-DNA interactions define the mechanistic aspects of circle formation and insertion reactions in IS2 transposition

Figure 3

Protein-DNA complexes visualized by gel retardation assays run on 5% polyacrylamide gels. For each lane, 80 nM of partially purified IS2 OrfAB-GFP was reacted with 2 nM 32P-labeled annealed oligonucleotides containing cognate DNA sequences from IRR, IRL, or the minicircle junction substrates MJcj and MJpc. The reactions were incubated at room temperature (20°C) for 30 min, loaded onto the gel at 4°C and run at 120 mA. (A) Lanes 1 to 3: 87-mer IRR; 4 to 5: 79-mer IRL. Different preparations of the protein were used in lanes 2 and 3. The gel was run for 1400 Vhr. (B) Lanes 1 and 2: 114-mer MJcj; 3 and 4: MJpc. The gel was run for 920 Vhr. IRR/IRL: right and left inverted repeats; MJcj: covalently joined minicircle junction substrate; MJpc: precleaved minicircle junction substrate.

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ISSN: 1759-8753