Preliminary Study On Transgenic Misgurnus Anguillicaudatus With PiggyBac

In order to explore transgenic fish research prospects of piggyBac transgenic system, we transported transposon vector pigA3GFP which carries the GFP gene into loach eggs proceded by artificial fertilization using sperm mediating together with the helper plasmid which encodes piggyBac transposase. In G0 generation, we obtained the loach emitting the green flurescence. The result of PCR and dot-blot hybridization experiment confirmed that the GFP gene has been integrated into loach genome.We constructed the transgenic vector pigA3GFP-Promoter-DsRed-PolyA based on pigA3GFP vector.The DeRed report gene was driven by Actin promoter from Megalobrama amblycephala and added the Poly(A) signal of the fibroin protein from Bombyx mori to its terminal sequence.The vector and auxiliary helper plasmid mixed was transferred into BmN cells through lipofection method.After the cells undergoers 72 hours cultured,we discovered the green and red fluorescence simultaneously in BmN cells. The result shows that the DsRed gene is able to be driven by the Actin promoter from Megalobrama amblycephala.