Figure 7.

TetR^S-dependent expression of p5R70Tet4mycMyoA and the compensatory modulation of the endogenous TgMyoA level. (A) Detection of mycMyoA by IFA, on intracellular parasites incubated in the presence or absence of ATc, using mAb anti-myc. (B) Analysis of mycMyoA and endogenous MyoA expression by western blot using anti-myc and anti-MyoA tail polyclonal antibodies. Detection of MIC4 with anti-MIC4 antibodies was included as reference. Before preparation of cell lysates, parasites were incubated for 48 h in the presence or absence of ATc. (C) Semi-quantitiative RT–PCR analysis using two dilutions of total RNAs isolated from wild type and parasites expressing myMyoA 48 h after incubation in the presence or absence of ATc. Specific sets of primers were chosen to amplify the endogenous MyoA, the transgenic mycMyoA and as control MIC4 transcripts.