Skip to main content
. Author manuscript; available in PMC: 2013 Nov 19.
Published in final edited form as: Mol Biochem Parasitol. 2005 Sep 6;144(2):10.1016/j.molbiopara.2005.08.009. doi: 10.1016/j.molbiopara.2005.08.009

Fig. 4.

Fig. 4

Double-inducible expression in cells with a hyg-tagged RRNA locus. (A and B) A reporter gene (GeneDB: Tb10.6k15.3240) was cloned in pT7aGFP (similar to pRPaGFP but with a T7 promoter in place of an RRNA promoter) and the nuclear GFP-tagged protein was expressed in T7i:Tag cells. (A) Induction was with 5ng ml−1 Tet for 24h. The western blot was probed with primary rabbit α-GFP (1:4000). The left hand panel shows a coomassie-stained gel as a loading control. (B) Direct detection of the GFP-tagged protein. DNA counterstained with (DAPI); k, kinetoplast; n, nucleus. (C) The p2T7aTAblue RNAi construct. Details as in Fig. 1A. (D and E) dsRNA against α/β-tubulin was expressed in T7i:Tag cells using p2T7aTUB, a derivative of p2T7aTAblue. Details as above. (D) Microscopy. Induction was with 5ng ml−1 Tet for 20h. (E) Growth analysis. Details as in Fig. 1D.