Figure 4

Analysis of 2,047 bp upstream of the Pb GP43 ORF. A, Size comparison of the PbGP43 5' flanking region from fourteen P. brasiliensis isolates. Ethidium bromide-stained agarose gel showing the amplicons produced with P4 (forward) and GRN (reverse) primers using genomic DNA from the indicated isolates. M, molecular markers. B, schematic representation of the PbGP43 5' flanking region from isolates Pb339/Pb18 and Pb3, where the positions of P4/GRN primers are shown. The repeated regions are boxed and start at the dark gray bar. The lighter-colored box indicates a 58-bp sequence ("connector", shown in C) that is absent in the upstream repeated region 1c and 1c/a/b. The sequences in the color-coded boxes can be found in the sites indicated in B by the correspondent colored arrow. D, sequence alignment of the Et12/Et23 probes (-255 to -215 in 1a region) with the correspondent fragments in other regions from Pb3, Pb18 and Pb339, as indicated. The overlap between these probes is indicated, as well as one of the connector sequences (brown) boxed in C.