Figure 8

Genomic organization of HSP70 genes from L. braziliensis. DNA from promastigotes was totally or partially digested with restriction enzymes and the resulting fragments were separated on a 0.8% agarose gel, transferred to nylon membrane and hybridized with the H70-IR-E probe (A). The same blot was stripped and rehybridized with the HSP70-3' UTR-II probe (B). Lanes: 1, 185 ng of λ DNA Hind III; 2, 300 ng of Xho I-digested DNA; 3, 1 μg of Xho I+Sma I-digested DNA; 4, 1 μg of Xho I+Bam HI-digested DNA; 5 to 9, DNA digested with Bam HI for 2 min (lane 5), 5 min (lane 6), 15 min (lane 7), 30 min (lane 8), and 3 hours (lane 9); lane 10, 140 ng of Φ29 DNA+Hin dIII. Molecular weight markers (lane 1 and 10) were labeled with digoxigenin and used as a probe in the hybridizations. (C) Pulsed field gel electrophoresis showing ethidium bromide staining of S. cerevisiae (lane 1), and L. braziliensis chromosomes (lane 2). Panel 3 shows the hybridization of the H70-IR-E probe to the L. braziliensis chromosomes. (D) Graphical representation of the deduced physical map for HSP70 locus in L. braziliensis. Small red boxes represent the 5' UTR; light blue boxes the 3' UTR-I, dark blue one the 3' UTR-II, and green boxes the ORFs.