In Pb339, identities between regions are 89% (1a × 1b), 79% (1a ×

In Pb339, identities between regions are 89% (1a × 1b), 79% (1a × 1c) and 90% (1b × 1c). In Pb18, the structure and sequence of the PbGP43 5′ flanking region (2,047 bp) are quite similar to those in Pb339. Sequence identities are also high when comparing the same regions between Pb339 and Pb18: 99% (1a), 95% (1b) and 97% (1c). Pb3 lacks one repetitive region: 1a in Pb3 is 96% identical to 1a in Pb339, while 1c/a/b carries nucleotides characteristic of the three regions, however the level of identity is higher with 1c (94%) than with 1b (87%) or 1a (78%). Therefore, when sequence alignments of the repetitive regions from

Pb339, Pb18 and Pb3 were compared in a dendrogram, there were two main clusters, one with 1a sequences and another branching into 1b and 1c (and 1c/a/b) regions (data not shown). Pb3 sequences formed individual Copanlisib research buy branches, in accordance with the phylogenetically distinct nature of this isolate detected with PbGP43 gene and other loci [3, 15]. The 442-bp upstream fragment is highly divergent from the repetitive Vistusertib regions, but conserved among isolates (about 99% identity). The highly conserved nature of the connector (Figure 4C) drove our attention to a more detailed analysis of its contents. We observed that some oligonucleotide sequences occur exclusively in the connectors, while others can be found

in other positions of the repetitive regions. In Figure 4C, we boxed six sequences (6- to 8-bp long) Doxacurium chloride that can be found in the positions represented in Figure 4B by color-coded arrowheads or bars. Note that the blue oligonucleotide (TTTTCAAG) was invariably found 44 bp upstream of the last base of all repetitive regions. The purple sequence (ATGAAAT) localized 109 bp downstream of the first base of the connector in the three isolates considered; therefore this sequence is not seen in 1c (or 1c/b/a) region. The gray sequence TTGATA in the connector could also be seen in 1b region at -883 (Pb339) and -1006 (Pb3). The green ATGTTA oligonucleotide was detected at -1756 (Pb339 and Pb18) and -1261

(Pb3) and at -268 in all isolates. The orange TATAGA was found exclusively in Pb18 and Pb339 at distances of 186 and 184 bp from the start base of 1a and 1b regions. The red-coded corresponding mutated sequence in Pb3 (TTATTGAT) was also detected 238 bp upstream of the last base in 1c/b/a region; it is not present in Pb18 or Pb339 connector, but it could be detected at distances varying among 237, 234 and 229 bp upstream of regions 1a, 1b and 1c last bases. The brown CTTATTT initial connector sequence was observed only once in 1a region, 67 bp upstream of the last base in Pb339 and Pb18. Although this exact sequence is not observed in the Pb3 connector, which shows a unique CTTCATT oligonucleotide not found elsewhere, in this isolate CTTATTT has been observed twice in 1a region, at 67 bp upstream of the last base, and at a polymorphic -372 site.

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