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. 2010 Apr 20;285(26):19767–19775. doi: 10.1074/jbc.M110.114520

FIGURE 5.

FIGURE 5.

Generation and characterization of L. infantum AdoMetT1 null mutant. A, genomic organization of the AdoMetT1 locus. The AdoMetT1 upstream and downstream fragments used for the integration of the NEO and ZEO cassettes are shown. P, PstI. UTR, untranslated region. B, Southern blots of L. infantum genomic DNA digested with PstI and hybridized to a AdoMetT1–3′ UTR probe. Lane 1, L. infantum wild type; lane 2, L. infantum with one AdoMetT1 allele disrupted with the NEO cassette; lane 3, L. infantum with one AdoMetT1 allele disrupted with a ZEO cassette; lane 4, L. infantum AdoMetT1 null mutant with integrated NEO and ZEO cassettes. C, sinefungin susceptibility of Leishmania cells. D, transport of 60 nm S-[3H]adenosylmethionine by Leishmania cells. ●, L. infantum wild type; ○, L. infantum AdoMetT1 with one inactivated allele; ▴, L. infantum AdoMetT1 null mutant; ▵, L. infantum AdoMetT1 null mutant transfected with a add-back AdoMetT1 plasmid. The average of triplicate measurements is shown for C and D.