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. 2010 May;9(5):784–794. doi: 10.1128/EC.00336-09

Fig. 6.

Fig. 6.

Construction of transgenic P. yoelii parasites to test for functional complementation of UIS4. (A) Strategy for replacing PyUIS4 with PF10_0164myc or PyUIS4myc in P. yoelii. The transgenes are under the control of 1.4 kb of the endogenous PyUIS4 promoter and the PbDHFR 3′ UTR. The PyUIS4 5′ UTR and 3′ UTR target the constructs to the endogenous PyUIS4 locus. The mutant TgDHFR/TS enzyme is used in conjunction with pyrimethamine to select for transgenic parasites. Genotyping primers anneal where indicated to test for the presence of the PF10_0164 ORF (Pf) or the PyUIS4 ORF (Py) or for integration of the construct into the genomic locus from the 5′ or 3′ end. The boundary between the thick and thin black bars in the “clone parasite” line indicates the transition from chromosomal DNA to the integrated construct. (B) PCR-based genotyping of Pyuis4(−)PF10_0164myc, PyUIS4myc, and PyWT parasite clones. Amplicons represent the presence of the PyUIS4 ORF (Py) and the PF10_0164 ORF (Pf) and integration of the relevant construct into the PyUIS4 genomic locus from the 5′ or 3′ end.