Schematic of pTb-AMluc constructs containing luciferase reporter genes.

<p>We used a modular format for the construction of the pTb-AMluc series of vectors that were designed to target the “red-shifted” luciferase genes <i>PpyRE9H</i> and <i>PpyRE-TS</i> (red) to the rDNA locus of <i>T.b. brucei</i>. The ribosomal locus targeting fragments (green), and <i>VSG</i>, <i>GPEET2 procyclin</i> and <i>tubulin</i> flanking sequences (blue) were derived from parasite DNA by PCR as outlined in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002571#s2" target="_blank">Materials and Methods</a> and Supplementary Table 1. IR refers to the intergenic region that the contains processing signal(s) at the 5′-end of the puromycin N-acetyl transferase (<i>PAC</i>) (purple) and the 3′-end of the <i>REH9</i> luciferase genes. Constructs were linearised by SacI/KpnI digestion prior to transfection.