单精注射法生产转基因小鼠的研究  

Production of Transgenic Mice Expressing GFP by Intracytoplasmic Sperm Injection(ICSI)

作　　者：苗向阳[1] 邵建军[1] 方昌阁[1] 丁淑燕[1] 任慧英[1] 朱瑞良[2] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所 [2]山东农业大学动物科技学院,泰安271018

出　　处：《畜牧兽医学报》2007年第6期548-551,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：中国农业科学院院长基金;创新基金资助项目(9725-16)

摘　　要：精子胞质内显微受精技术(ICSI)作为辅助受精的一种手段,将体外受精研究和胚胎的显微操作技术结合起来,比以往对精子质量的要求大大降低,使其无论在畜牧业生产实践还是在哺乳动物的生殖生理基础研究中都有着十分重要的意义。本研究用小鼠精子及小鼠精子与GFP基因孵育后对小鼠卵母细胞进行ICSI,获得子代鼠。提取鼠尾基因组DNA,应用PCR、Southern blot进行整合检测。在发育至成年的11只小鼠中经PCR和Southernblot检测到3只阳性(27.3%)。结果表明,利用ICSI技术可以高效地生产转基因小鼠。Intracytoplasmic sperm injection （ICSI） is the process in which a high powered micro- scope is used to inject specially prepared single spermatozoa into a mature oocyte in order to effect fertilization. ICSI is now considered to be a useful procedure, not only for producing live offspring from nonmotile sperm cells but also for generating transgenic animals through sperm-mediated gene transfer. Mouse ICSI was conducted using mouse sperm or sperm which were incubated with GFP gene for 30 minutes. Genomic DNA was extracted from the offspring of the founder mice for PCR and Southern blotting analysis. PCR and Southern blotting analysis showed 3 of 11 mice were transgenic. These results indicated that the high efficiency of transgenic mouse could be produced by ICSI.

关 键 词：单精注射 转基因 胚胎 

分 类 号：S813.3[农业科学—畜牧学]