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. 2016 Jun 17;12(6):e1006117. doi: 10.1371/journal.pgen.1006117

Fig 1. RAD50 gene conditional inactivation in L. infantum.

Fig 1

(A) Schematic representation of the RAD50 locus in L. infantum before and after integration of the inactivation cassettes blasticidin-S deaminase (5’-BLAST-3’), puromycin acetyltransferase (5’-PURO-3’) and transfection construct Psp-NEO-RAD50. S, SacI restriction sites. (B, C) Southern blot analysis with genomic DNAs digested with SacI were hybridized with probes covering either the 5’ flanking region of RAD50 (B) or the RAD50 ORF (C). (D) PCR analysis with primers set aa’ and bb’ the chromosomal copies of the MRE11 and RAD50 genes respectively. Lanes: 1, L.infantum WT; 2, WT Psp-NEO-RAD50; 3, WT Rev Psp-NEO-RAD50; 4, RAD50-/- Psp-NEO-RAD50; 5, RAD50-/- Rev Psp-NEO-RAD50 grown for 55 passages in absence of G418; 6, MRE11-/- and 7, MRE11-/-RAD50-/-.