Fig. 1. : restriction maps and genetic elements of the Gateway® plasmids (A) pTEX-TAPtag-GW map. The ampicillin resistance gene (neo r), Gateway® cassette and TAPtag sequence are flanked by the 5’-upstream and 3-downstream regions of the Trypanosoma cruzi glycosomal glycosomal glyceraldehyde-3-phosphate dehydrogenase gene (gGAPDH) genes. The Gateway® cassette consists of the attR1 and R2 sites (black boxes), the chloramphenicol resistance gene (Cmr) and the ccdB gene. The TAPtag cassette consists of the calmodulin-binding peptide (CBP), protein A (PA) and a spacer region with a Tobacco Etch Virus (TEV) protease cleavage site; B: pTREX-TAPtag-GW map. This vector has the same features as pTEX with the addition of a ribosomal promoter (with a transcription starting point, black arrow head) and the HX1 fragment; C: inducible expression vector pTcINDEX-GW. The grey box indicates the HX1 fragment. The T. cruzi actin intergenic region (TcActin IG), the T7 transcriptional terminator (T) and the Gateway® cassette are shown. The black flags indicate the T7 promoter and the grey circle denotes the location of the tetracycline operator. R-NTS/P is the ribosomal non-transcribed spacer and promoter region used for targeting. The Roman numerals I and II indicate the two halves of the targeting sequence cloned in the opposite orientation of that in the genome. The black arrowhead indicates the location of the polymerase I transcription start site.