FIG. 5.

RT-PCR analysis of C. parvum TRAPC1 expression during in vitro development in HCT-8 cells. (A) Total RNA (2.0 μg) isolated from C. parvum-infected HCT-8 cells at 6, 12, 24, 48, and 72 h p.i. (lanes 1 to 5, respectively) was reverse transcribed by using random primers, and 2.0 μl of a 1:50 dilution of each cDNA reaction was subjected to 23 cycles of amplification in the presence of [³²P]dCTP and primers specific for C. parvum 18S rRNA (Table 1). P, positive control (C. parvum genomic DNA used as a template in PCR); N, negative control (total RNA from mock-infected cells used in RT). (B) Two microliters of each cDNA reaction was amplified by using primers specific for C. parvum TRAPC1 (Table 1).