*2.11. Construction of HA-Tagged EhTrxR Trophozoites*

For the construction of the pJST4-EhTrxR expression vector that was used to express HA-tagged EhTrxR in the parasite, EhTrxR was amplified from *E. histolytica*'s genomic DNA using the primers 5- EhTrxR\_KpnI (ggtaccatgagtaatattcatgatg) and 3- EhTrxR\_BamHI (ggatccatgagtttgaagcc). The resulting PCR product was cloned into the pGEM-T Easy vector system (Promega, WI, USA) and then digested with the restriction enzymes, KpnI and BamHI. The digested DNA insert was subcloned into the *E. histolytica* expression vector pJST4, which was previously linearized with KpnI and BamHI. The pJST4 expression vector contains a tandem affinity purification tag for use in protein purification and identification [35]. This CHH tag contains the calmodulin binding protein, hemagglutinin (HA), and histidine (His) residues, and its expression is driven by an actin promoter.
