Highly Sensitive In Vivo Imaging of Trypanosoma brucei Expressing “Red-Shifted” Luciferase
Figure 1
Schematic of pTb-AMluc constructs containing luciferase reporter genes.
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 PpyRE9H and PpyRE-TS (red) to the rDNA locus of T.b. brucei. The ribosomal locus targeting fragments (green), and VSG, GPEET2 procyclin and tubulin flanking sequences (blue) were derived from parasite DNA by PCR as outlined in Materials and Methods 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 (PAC) (purple) and the 3′-end of the REH9 luciferase genes. Constructs were linearised by SacI/KpnI digestion prior to transfection.