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将稀有鲫 (Gobiocyprisrarus)精子与重组质粒pCAhLFc线性DNA混合温育 ,经电脉冲处理后与卵子受精 ,孵化出苗。从鱼苗中提取DNA ,经PCR检测 ,2 5 .5 %～ 6 6 .7%鱼苗带有外源基因。在显微镜下观察经电脉冲处理过的精子 ,发现其活力有不同程度下降 ,受精率也有不同程度下降 ,说明不同的电脉冲条件对精子有不同程度的损害作用。精子与外源DNA混合温育 ,经电脉冲处理后 ,用DNA外切酶消化后 ,提取精子DNA ,经PCR检测 ,仍有阳性电泳带 ,证明电脉冲可以促使稀有鲫精子摄入外源基因
The gobiocyprisrarus sperm and recombinant plasmid pCAhLFc linear DNA mixed incubation, after the electric pulse treatment and fertilization of eggs, hatching emergence. DNA was extracted from fry and detected by PCR. 25.5% -6.67% of the fry had foreign genes. Under the microscope, the electrical pulse-treated spermatozoa were found to have different degrees of decline in vitality and the fertilization rate decreased to some extent, indicating that different electrical pulse conditions have different degrees of damage to sperm. Sperm and exogenous DNA mixed incubation, after the electric pulse treatment, digested with DNA exonuclease digestion, sperm DNA, detected by PCR, there are still positive electrophoresis zone, that electric pulse can promote the absorption of sperm into the exogenous gene