Figure 2

Genomic organization of L. braziliensis α-tubulin genes. Southern blot of promastigote DNA hybridized with the α-tubulin ORF. (A) Two μg of L. braziliensis DNA were partially digested with Csp451 for 2 min (lane 2); 5 min (lane 3) and 15 min (lane 4); lane 5 contains 1 μg of totally digested DNA. Thin arrows in panel A point to the ladder bands; a thick arrow (in panels A and B) points to the repeated unit; the arrow head and the asterisk (in panels A and B) mark additional hybridization bands. (B) Southern blot corresponding to 3 μg of L. braziliensis DNA digested with either PstI (lane 2) or ApoI (lane 3); 5 min of film exposure. Lane 4 shows a Southern blot of 3 μg of Csp451-digested DNA; 20 min of film exposure. Thin arrows point to hybridization bands. Molecular weight markers: lanes 1, a mixture of undigested and HindIII-digested DNA from λ phage; lanes 6, 1-Kb plus marker (Invitrogen California, USA). The position of the DNA markers in the blot was revealed by including digoxigenin-labeled markers in the hybridization mixture. As probe, the L. braziliensis α-tubulin ORF was used. (C) Hypothetical map for the α-tubulin locus in L. braziliensis chromosome 13, as deduced from Southern blot analyses and genomic sequence at GeneDB database (D). Map for α-tubulin locus in L. braziliensis chromosome 29, as deduced from genomic sequence at GeneDB database. Pentagonal shaped boxes represent α-tubulin genes, numbers inside each box are verb GeneDB entries (LbrM.13.0190, LbrM.13.0200, LbrM.13.0210 and LbrM.29.2700); filled boxes at the end of each gene represent the 3´ UTR; and rectangles located at the 5´ end of each gene, the 5´ UTR.