Generation of the <i>L</i>. <i>infantum chagasi</i> line expressing luciferase (Lc-LUC).

<p>(<b>A</b>) Schematic representation of the SSU rDNA locus after integration of the linearized construct derived from <i>pSSUint-Luc2</i>. SSU: small subunit; SAS: splice acceptor site; LUC2: <i>luc2P</i> coding sequence; <i>αTub</i> IR: intergenic region of the <i>L</i>. <i>enrietii α-tubulin</i> gene (El Fadili et al., 2002); HYG: hygromycin phosphotransferase gene; <i>CPB-2</i>.<i>8</i> IR: <i>L</i>. <i>mexicana CPB 2</i>.<i>8</i> intergenic region. The hatched bar indicates the transfected cassette. Position of primers and size of amplified fragments are indicated by dotted lines. (<b>B</b>) Size separation of PCR products obtained from Lc-WT (lanes 1, 3, 5 and 7) and Lc-LUC (lanes 2, 4, 6 and 8) genomic DNA with primers S1 and S4 (1 and 2), S1 and Luc2Rev (3 and 4), Luc465–484 and S4 (5 and 6), Luc2For and Luc2Rev (7 and 8). (<b>C</b>) Correlation between luciferase activity and number of Lc-LUC promastigotes. Parasites were serially diluted and luminescence was measured using a microplate reader. Results are the mean and standard deviation of triplicate determinations from a representative experiment. RLU: relative luminescence units.