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. 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. 2.

Fig. 2

Tagging an RRNA locus. (A) NotI-digested pRPGFP and SalI/SacI-digested ph3E were used to identify an RRNA locus with no evidence of position effect repression and to add the hyg-tag to that locus respectively. The resultant tag can then be replaced using AscI-digested pRPaGFP. HYG, hygromycin resistance; PAC, puromycin resistance; ACT, actin; ES, variant surface glycoprotein Expression-Site. (B) Western blot analysis of inducible expression in three independent clones generated using non-tagged cells and pRPcMyc with a reporter gene (GeneDB: Tb927.4.2520). The blot was probed with primary mouse α-cMyc (1:2000). (C) Western blot analysis of induced expression in three independent and representative clones generated using tagged RRNA locus cells and pRPaGFP with a reporter gene (GeneDB: Tb11.01.3380). Lane 1: un-induced cell line 1. The blot was probed with primary rabbit α-GFP (1:4000). Other details as in Fig. 1.