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As an effective method of transgenesis, the plasmid of  PiggyBac  transposon containing GFP ( PiggyBac ) transposon system has been widely used in various organisms but not yet in mollusks. In this work,  piggyBac  containing green fluorescent protein (GFP) was transferred into the Pacific oyster ( Crassostrea gigas ) by sperm-mediated gene transfer with or without electroporation. Fluorescent larvae were then observed and isolated under an inverted fluorescence microscope, and insertion of  piggyBac  was tested by polymerase chain reaction (PCR) using genomic DNA as template. Oyster larvae with green fluorescence were observed after transgenesis with or without electroporation, but electroporation increased the efficiency of sperm-mediated transgenesis. Subsequently, the recombinant  piggyBac  plasmid containing  gGH  ( piggyBac-gGH ) containing GFP and a growth hormone gene from orange-spotted grouper ( gGH ) was transferred into oysters using sperm mediation with electroporation, and fluorescent larvae were observed and isolated. The insertion of  piggyBac-gGH  was tested by PCR and genome walking analysis. PCR analysis indicated that  piggyBac-gGH  was transferred into oyster larvae; genome walking analysis further showed the detailed location where  piggyBac-gGH  was inserted in the oyster genome. This is the first time that  piggyBac  transposon-mediated transgenesis has been applied in mollusks.

PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks