A fast-growing trait of loach (Misgurnus anguillicaudatus) obtained by sperm-mediated gene transfer biotechnology

Abstract

An antifreeze protein gene promoter from ocean pout Macrozoarces americanus linked to a chinook salmon Oncorhynchus tschawytscha growth hormone cDNA (opAFPGHc) was introduced into loach Misgurnus anguillicaudatus sperm via electroporation. Compared to the untreated sperm, the fertilization rate driven by the electroporated-sperm decreased slightly, but had not significant effect on hatchability. The success rate of gene transfer was 50%. Genomic Southern blotting demonstrated that DNA from transgenic loach contained positive band(s) with higher (37.5%), the same, or lower molecular size than the transferred opAFPGHc fragment. There was a mosaic distribution of exogenous DNA in a wide variety of tissues analyzed. The average weight gain between the 14th and 161th day of the experimental group was 2.5-fold higher than that of the control and the mock-treated groups. The body length distribution of the experimental group was rather widely scattered instead of being normally distributed.