FIG 2 .

Transcription and expression of Pvs48/45 in transgenic parasites. (A) PCR detection using cDNAs prepared by reverse transcription using universal oligo(dT)18 primer. VL, primer pairs (VL-F and VL-R) for a long product (1,214 bp) from pvs48/45; VS, primer pairs (VS-F and VS-R) for a short product (452 bp) from pvs48/45; BS, primer pairs (BS-F and BS-R) for a short product (423 bp) from pbs48/45 were used for PCR amplification. The PCR results from WT, Tr-F, and Tr-C parasites are shown in the left panel, while the results from KO parasite are shown in the middle panel. The quality of the cDNAs obtained from all the parasites was tested using primers for an unrelated gene (pbdmc1) as a positive control (right panel). (B) (Left panel) PCR detection using cDNAs prepared by reverse transcription using gene-specific primers. (Right panel) PCR results obtained using DNase I-treated RNA samples without reverse transcription. The use of the assay ruled out the possibility of genomic DNA contamination. (C) The gametocyte-enriched parasite proteins of WT, KO, Tr-F, and Tr-C were fractionated by SDS-PAGE, transferred to nitrocellulose membranes, and probed with anti-Pvs48/45 mouse serum samples (upper panel) and anti-Pbs48/45 rabbit serum samples (middle panel). Anti-PfHSP70 mouse serum samples were used as a control for confirmation of the quality of the parasite lysates (lower panel).