Fig. 4

RT-PCR detection of LUC mRNAs in T. cruzi stable transfectants and of the 5′ end nature of Luc mRNAs. Cytoplasmic RNA from T. cruzi stable transfectants eletroporated with pTEXPr77Luc or pTEXPr77Mut1-17Luc were used as templates for reverse transcription and subsequent PCR amplification of the Luc mRNA 5′ end using a primer corresponding to the Pr77 5′ end (Pr77-LUC), or the SL primer (SL-LUC), as sense primers. As a control of RNA quality, the 5′ end of the kmp11 RNA was reverse transcribed and PCR amplified. The pTEXPr77Luc and pGEMTSL-Luc plasmids were used as DNAs template in positive control reactions; no DNA was used in negative control reactions. Mutants are numbered from 1 to 17 (M1-17)