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. Author manuscript; available in PMC: 2013 Nov 1.
Published in final edited form as: Mol Biochem Parasitol. 2012 Aug 28;186(1):11–20. doi: 10.1016/j.molbiopara.2012.08.005

Figure 2. L. major pla2− mutants grow normally as promastigotes in culture.

Figure 2

(A–B) Southern blot showing the replacement of PLA2/PAF-AH alleles with marker genes (PAC/BSD) in pla2−. Genomic DNAs from WT, PLA2+/− (heterologous parasites in which one of the two PLA2/PAF AH alleles has been deleted), and pla2− promastigotes were digested with BglII and HindIII and resolved on an agarose gel. Following transfer to a PVDF membrane, a radiolabeled DNA probe was used to detect the PLA2/PAF-AH allele (~3.3 Kb) and the PAC/BSD replacement alleles (10.5–10.7 Kb). (A) Result from autoradiography. The asterisk represents a non-specific band recognized by the probe. For pla2−, BSD and PAC replacements appear as one band due to the similarity in size. (B) Schematic representation of the Southern blot. (C) Promastigotes were inoculated in M199 medium and culture density was determined daily using a hemacytometer. Error bars represent standard deviations from 2 experiments.