抄録
We used loach Misgurnus anguillicaudatus sperm to study the effect of the electroporation conditions for fish sperm on the capacity of transferring a foreign gene into the eggs and the development of their derivative embryos. Results showed that sperm electroporated at low field strengths resulted in a higher hatching rate and a lower abnormality rate, compared to the high field strength group. However, the success rate of gene transfer of the low field strength group was lower thanthat of the high field strength group. Sperm electroporated at a concentration of 175 μg/ml plasmid DNA had a higher gene transfer rate than did those at 100 μg/ml or lower. The hatching rate of embryosderived from ova fertilized with sperm treated at 250 μg/ml DNA decreased abruptly, even at a low field strength (3.5 kV). Increasing the number of pulses per cycle (up to 211) and the number of cycles (upto 12) enhanced the success rate of gene transfer. Dot and Southern blot analyses and PCR detection proved the existence of transferred DNA in embryos following sperm-electroporation. An optimal conditionof electroporation for loach sperm to obtain normal development in transgenic loach is suggested.
