转抗冻蛋白基因鱼的研究

Transgenic Goldfish of Antifreeze Protein Gene

鱼类抗冻蛋白(AFP)或抗冻糖蛋白(AFGP),可使鱼类体液的冰点降低至-1.9℃左右,并减少冰晶的生长速度,它们的生物活性比小分子NaCl等无机盐、尿素。

New foundland ocean pout antifreeze protein (AFP) gene after removing negative regulation sequence were microinjected into goldfish fertilized eggs. Individual fingerlings were analyzed for AFP gene by Dot hybridization and Southern blotting. DNA from 3 out of 47 in experiment 1 and from 4 out of 25 fingerlings in experiment 2 showed Dot hibridization to the AFP gene. Southern analysis of DNAs from 2 positive Dot hybridization showed also all positive hybridization bands to the AFP gene probe. After PCR amplification of the DNAs from 4 positive Dot hybridization, there were 3 positive DNA Southern hybridization samples. These hybridization signals were absent in the DNA from control fish.We discussed also here the methodology of incresing positive frequency and expression level of transgenic fish.