Exploration of transgenic method for goldfish and loach with the piggyBac transposon

Abstract

Development of transgenic fish techniques and application will promote traits improvement of fish and benefit exploration of its gene function.To investigate the applicability of the transposon piggyBac in Cyprinidae and Cobitidae fish,the expression element of DsRed under the control of PAβ promoter was inserted into the plasmid pigA3GFP to generate transgenic vector pigA3GFP-AβDsRed.The results of the CIK cell transfected with pigA3GFP-AβDsRed showed that the transposon piggyBac could transfer exogenous genes into CIK genome,and both of the promoter A3 from silkworm and the promoter CMV from CMV have the bioactivity.Moreover,the pigA3GFP-AβDsRed and pigA3GFP were mixed with helper plasmid helper-pigA3 respectively,and then were introduced into the eggs of goldfish and loach by sperm-mediated transfer.The results of fluorescence observation,PCR and Dot blotting verified that both transgenic goldfish and loach were obtained.Therefore,PB transposon was considered to be active in some fishes of Cyprinidae and Cobitidae.