Study On The Two Approaches About Gene Transfer Methods To Turbot (Scophthatmus Maximus)

Comparative study on the gene transfer into Turbot (scophthatmus maximus) zygotesby electroporation and transfection were conducted. The objective gene waselectroporated and transfected into turbot eggs with sperm.Based on fish hatching rate and gene transfer rate,single-factor experiments weredesigned to optimize the parameters for the integration of foreign DNA into theturbot sperm cells during electroporation process. The electroporation condition(field strength of 300 V/cm, 5 pluse and foreign DNA concentration of 20mg/L) wasapplied for mass gene transfer. Procedures of preparing jetPEI/sodium chloridemixtures followed the vendor's manual (Qbiogene). Transfection experiments wereperformed at the one-cell stage (within 50 min postspawning) of fertilized fish eggs.Efficiency rates of foreign gene in the mass gene transfer were about 20~28% and60~70% for electroporation and transfection methods, respectively. It indicated thattreating the fish zygotes with the DNA/jetPEI complex at the one-cell stageexhibited higher gene transfer efficiency in fish.The preliminary study on electroporation and transfection were proved to be validways for transgene and its application to turbot.