Figure 4

Validation of EhSINE 2 polymorphic loci 18 and 50. PCR was performed using genomic DNA of E. histolytica HM-1:IMSS (H) and Rahman (R) as template, using primers from sequences flanking the EhSINE2 copy (number as shown on top). The size of amplicons was determined by electrophoresis in 1% agarose gel. EhSINE2 was missing in Rahman at the two loci, as the amplicon from Rahman was shorter by ~700 bp (the size of EhSINE2) at these loci (Panel (i)). The sizes of amplicons obtained are indicated on the right (arrows). The absence of EhSINE2 was further confirmed by Southern blotting with EhSINE2 probe, which failed to hybridize with the amplicons of strain Rahman (Panel (ii)). The specificity of the amplicon in Rahman was checked by Southern blotting with locus specific probe (Panel (iii)), which hybridized with the amplicons in both strains.