α-Actinin TvACTN3 of Trichomonas vaginalis Is an RNA-Binding Protein That Could Participate in Its Posttranscriptional Iron Regulatory Mechanism
Figure 6
Expression and recognition of TvACTN3 domains DIr, DIIr, and DIIIr by α-TvACTN3r antibody. (a) Map of domains used to generate the recombinant proteins (DIr, DIIr, and DIIIr) of TvACTN3. ((b)–(d)) Induction (I), purification (P), and recognition by WB of DIr, DIIr, and DIIIr proteins. Expression of the 6x-His tagged DIr, DIIr, and DIIIr proteins. Bacteria E. coli were transformed with pProEX-HTb- DIr, DIIr, or DIIIr plasmid and protein expression was induced by the addition of 1 mM IPTG for 16 h at 16°C. Protein extracts were separated through 10% SDS-PAGE and gels were CBB-stained, IPTG-induced (I) bacterial extract (lanes 1). Affinity purified recombinant proteins using Ni-NTA-Sepharose (lanes 2; P). Immunodetection of DI, DII, or DIII polypeptide by WB assays using α-TvACTN3r serum (lanes 3). kDa, molecular weight markers in kilodaltons (Bio-Rad).
