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. 2012 Aug 9;40(19):9903–9916. doi: 10.1093/nar/gks735

Figure 2.

Figure 2.

PfSR1 is a functional SR protein that can mediate pre-mRNA splicing. Recombinant PfSR1 can complement S100 extracts and rescue the splicing activity similar to SRSF1. (A) Schematic of the expression vector used to express the recombinant PfSR1 and SRSF1 in 293 T cells. cDNA of both genes was cloned into the expression vector fused to a His-tag. (B) SDS–PAGE presenting the purified SRSF1 and PfSR1 loaded on gel in increasing volumes (1, 2 and 4 µl, respectively). (C) Complementation of splicing activity of PfSR1 and SRSF1 in S100 extracts (in vitro splicing assay). Increasing amounts of purified SRSF1 or PfSR1 were added to S100 extracts in the presence of radiolabeled Ftz reporter pre-mRNA. Lane 1: labeled Ftz pre-mRNA only; Lane 2: nuclear extract (NE) + Ftz; Lane 3: S100 extract + Ftz; Lane 4: NE + S100 + Ftz; Lanes 5–6: S100 + Ftz with increasing amounts of SRSF1; Lanes 7 and 8: S100 + Ftz with increasing amounts of PfSR1. Schematics of pre-mRNA, spliced mRNA and the spliced intron are shown in the left.