Figure 3
From: Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5
Generation of Licpa null mutants. A) Pst I restriction map of ΔLicpa C1, ΔLicpa C2 and ΔLicpa C1::CPA loci. The size of the restriction fragments expected is indicated. 5'FR and 3'FR indicate the 5' and 3' LiCPA flank sequence, respectively. The arrowheads indicate the position of the different primers used to confirm the correct integration of the disruption constructs by PCR. B) Southern blot analysis of the ΔLicpa C1 mutant DNA digested with Pst I and hybridized with the following probes : LiCPA 3'FR (lane A), ORF (lane B) and 5'FR (lane C). Lane D shows L. infantum DNA hybridized with the 3'FR probe. C) Southern blot analysis of ΔLicpa C2 mutant (lane A) and wild type (lane B) DNA hybridized with the LiCPA 5'FR probe. The single and double arrowheads indicate the deleted and the wild-type alleles, respectively.