Figure 4

In vitro translation and binding of the 2488 putative repressor protein. (A) SDS-PAGE of in vitro translated and FluoroTect™Green labelled proteins: a negative control reaction containing no DNA template (no DNA), a positive control reaction for the expression of luciferase (Luc), and the expression of 2488prp (pMPF2488 as DNA template). A prestained protein ladder (Fermentas) was applied. (B) EMSA using 60 ng of in vitro translated, unlabelled 2488prp (see Fig. 4A). Then, 15 ng labelled oligonucleotides, one covering the respective area of the xyn2 promoter (Lpxyn2, see Table 1), and another bearing a mutation from AGAA to CTCC (Lpxyn2 M, see Table 1) were applied without protein sample (lanes 1, 2). In vitro translated Xyr1 was used as a positive control.