StarGate® Fusion Cloning System example


Fig.1 shows a bicistronic Destination Vector that consists of two genes, PR65 and GFP, cloned into 2 separate Donor Vectors that were fused by means of pNFUSE-IBA-IRES1. The insert of the resulting Fusion Donor Vector was transferred in a second step into pESG-IBAwt1. Transcription of both genes in the bicistronic arrangement is controlled by the CMV promoter. While translation of upstream PR65 is cap-dependent, separate translation of downstream GFP is mediated by the intergenic IRES sequence.
Since transfected cells are fluorescing green they can be detected easily and may be selected and enriched to optimize yields of the coexpressed PR65 target protein (Fig.2).