Figure 7 | Scientific Reports

Figure 7

From: An efficient strategy for producing a stable, replaceable, highly efficient transgene expression system in silkworm, Bombyx mori

Figure 7

Strategy for site-specific gene integration via phiC31-mediated RMCE and subsequent marker excision from the target locus using the FLP/FRT system.

Production and selection of transgenic silkworm strains contained a single copy of the target gene 1 (Traget 1) without any transposon and marker genes (Marker 1 and Marker 2) in their genomes (Target loci) using the piggyBac-derived plasmid and as described in Methods and shown in Figure 1. Recombination between two attB/attP pairs from donor vector and the target loci mediated by phiC31 integrase (Int) resulted in integration of a single copy of the taget gene 2 (Target 2), the Marker 1 and an inducible promoter (IP)-driving FLP recombinase expression cassette (Ter indicates the termination sequence) into the target loci of the silkworm genome. Subsequent inducible expression of FLP recombinase (FLP) in vivo, which can catalyze recombination between two FRT sites and resulting excision of the Marker 1 and the FLP recombinase expression cassette from the target loci of the silkworm genome.

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