电脉冲作用将外源基因导入稀有鮈鲫精子的研究  

Introduction of Foreign DNA into the Genome of Rare Minnow by Sperm Electroporation

作　　者：钟家玉[1] 茅卫锋[1] 朱作言[1] 

机构地区：[1]中国科学院水生生物研究所淡水生态与生物技术国家重点实验室,武汉430072

出　　处：《Acta Genetica Sinica》2002年第2期128-132,共5页

基　　金：<国家重点基础研究发展规划>项目G2 0 0 0 0 1610 9资助

摘　　要：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 。Gene transfer was investigated in rare minnow via electroporated sperm. The sperm of the fish mixed with linear DNA (pCAhLFc) was electroporated. Then mature eggs were in vitro fertilized with these sperm cells. The DNA was extracted from the fries and analyzed by PCR. The percentage of fries with foreign genes varied between 25.5% and 66.7%. The observation of the electroporated sperm under microscope indicates that both the vigor of sperms and the successful ratio of the fertilization diminish, indicating that the electroporation with various parameters can hurt the sperm on different extent. The sperm cells electroporated with foreign genes, then was treated by DNA, and the DNA was extracted and was analyzed by PCR. It was found that foreign gene still exists, which approves that the electroporation can force the foreign gene enter the sperm of rare minnow.

关 键 词：稀有ju鲫 电脉冲-精子介导转基因法 外源基因导入 转基因鱼 

分 类 号：S961.6[农业科学—水产养殖]