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. 1998 Jul 21;95(15):8526–8531. doi: 10.1073/pnas.95.15.8526

Figure 4.

Figure 4

Binding of TnpD to DNA and DNA–TnpA complexes. A 279-bp DNA fragment containing two TnpA binding sites was used as the binding target DNA (Fig. 1b). Binding reactions were performed in DNA binding buffer and contained 80 ng of TnpA and 10 μg of protein from cell extracts. The extra, slow-migrating band in lanes 8 and 12 is likely the result of nonspecific interaction of proteins (BSA in lane 8 and NT1 cell extract in lane 12) with TnpA bound to DNA at the high protein concentrations used to equalize the total amount of protein. DNA–protein complexes were fractionated on a native 4% polyacrylamide gel. TnpA, E. coli over-expressed TnpA protein; TnpD(CE), extract of tobacco SR1 cells expressing TnpD; NT1(CE), extract of tobacco NT1 cells; Sp. comp., specific competitor DNA; Non-sp. comp., non-specific competitor DNA.