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. 2009 Oct 30;285(1):453–463. doi: 10.1074/jbc.M109.048850

FIGURE 7.

FIGURE 7.

A, Southern blot of genetically manipulated parasites. Genomic DNA was extracted from the following cell lines: wild type parasites (WT), heterozygous parasites (HYG/DHS34), heterozygous parasites transfected with pX63PAC-DHS34 (HYG/DHS34[DHS34]), and six clonal cell lines (clones 1, 3, 4, 7, 8, and 11) derived from the HYG/DHS34[DHS34] strain transfected with the PHLEOΔdhs34 gene replacement construct. B, the top panel shows a map of the DHS34 genomic locus and the locations of the primers used for PCR analysis. Genomic DNA from HYG/DHS34 heterozygous parasites (first panel) and from clones 2 and 4 was used as a template for PCR analysis. Primers 2 and 4 were designed to match the upstream region of the DHS34 gene, with primer 4 corresponding to a region (5′F) that was present in the gene deletion construct used for the chromosomal DHS34 replacements. Primer 1 was designed as a forward primer corresponding to the early coding region. Primers 5–10 were reverse primers corresponding to various regions of the DHS34 coding region. Molecular size markers (in kb) are indicated on the left.