Both constructs were able to direct translation of the EGFP cistron effectively as expected (Figure 2a and Figure2d). The use of an authentic EMCV IRES to direct translation of the lacZ cistron was also effective (Figure 2e), with levels of b-galactosidase activity comparable to those of the b-galactosidase reporter ( Figure 2f ). However, no b-galactosidase activity was detected from cells transfected with the pEGFP-C1/JV59GS/lacZ construct (Figure 2b ), demonstrating that the JV 59GS was unable to initiate translation, and therefore, in this context, did not possess any IRES-like functions.