Application of asymmetric burst pulse system for preparation of transgenic medaka (Oryzias latipes)

We have assembled the asymmetric burst pulse system that can generate a single rectangular high voltage pulse in nanosecond range and some low voltage burst pulses with both polarities between microsecond and millisecond ranges. It was able to introduce some chemical substances into fertilized medaka eggs easily and efficiently with this pulse system. As a next challenge, a preparation of a transgenic medaka (Oryzias latipes) with the asymmetric burst pulse system was studied. Green Fluorescent Protein (GFP) plasmid vector, pEGFP-N3, was tried to introduce into fertilized medaka eggs by this system. GFP expression was observed in embryo at some pulse parameters by this system. These results indicated that plasmid vector was transferred to medaka embryo cell and expressed coding GFP successfully. 12.5% of transgenic medaka were achieved when H.V. short pulse of 50 ns with both pre- and post- burst pulses were used to introduce GFP.