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Fig. 2 | Parasites & Vectors

Fig. 2

From: Genetic validation of Leishmania genes essential for amastigote survival in vivo using N-myristoyltransferase as a model

Fig. 2

PCR analysis of NMT complemented double replacement HYG/PAC parasites. a Diagram of NMT locus and targeted single alleles containing replacement by either the HYG or PAC genes. 5′flank and 3′flank boxes represent NMT flanking regions used for gene targeting. 5′DHFR and 3′DHFR boxes represent 5′ and 3′ dihydrofolate reductase flanking regions. Arrows show positions of primers used in PCR analysis of clones in b–d. Lane 1: LV9 NMT HYG/PAC [NMT-TK]; Lane 2: LV9 NMTHYG/PAC [NMT-TK] clone 12; Lane 3: LV9 NMTHYG/PAC [NMT-TK] clone 18; Lane 4: LV9 NMTHYG/+; Lane 5: LV9 wild type; Lane 6: no DNA control. bPAC integration, expected band 2513 bp. cHYG integration, expected band 3488 bp. d Genomic NMT, expected band 2097 bp. Allelic NMT was only replaced completely for clone 12 while clone 18 retained the gene. Clone 12 was used to generate LV9 NMTHYG/PAC [NMT-TK][NMT-tdtom] or LV9 NMTHYG/PAC [NMT-TK][tdtom]

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