中文摘要：

在过去的近30年中，转基因技术在哺乳动物基因表达方面研究的应用已经成为实验生物学及应用生物学领域最为显著的进展之一．传统的制作转基因动物方法有显微注射法、逆转录病毒感染法和胚胎干细胞法等，但每种方法都有其缺陷，限制了其在今后转基因动物研究中的广泛应用．对小鼠体内生殖细胞进行外源基因转染，研究精子形成过程中制作转基因小鼠的效率．首先运用睾丸注射法将被脂质体包裹的绿色荧光蛋白表达载体（pIRES2-EGFP）注射到公鼠睾丸及附睾内，然后根据精子形成的不同阶段，分别于注射后7、16、30和42天与发情母鼠合笼，利用PCR和DNA印迹方法对新生小鼠进行基因组DNA检测．在各阶段所得新生小鼠中PCR阳性率分别为6．82％、0、56．86％和42．86％，DNA印迹检测阳性率分别为6．82％、0、47．06％、34．69％．经活体荧光成像系统及荧光显微镜分析，转基因小鼠呈现绿色荧光表达．通过比较精子生成各阶段转基因效率高低，为以后通过用睾丸内注射法转染雄性牛殖细胞高效制作转基因动物提供了理论依据．

英文摘要：

In the past 30 years, the research and application of transgenic technology in gene expression of animal had become a noticeable advancement in experimental biology and applied biology. Microinjection, retrovirus mediated method and embryonic stem cells method were all the traditional methods of producing transgenic animals. But these methods had defects and limited application in the research of transgenic animals in the future. Exogenous gene was transferred into germ cells of male mice, in order to study efficiency of transgenic mice in different spermiogenesis stages. The green fluorescence protein expression plasmid（plRES2-EGFP） and liposomes were mixed and injected into male mouse testis and epididymis. Then the intratesticular mice were mated with female at day 7, 16, 30 and 42 after infection. Polymerase chain reaction（PCR） and Southern blot were applied to identify transgenic mice. The positive ratio was 6.82%, 0, 56.86%, 42.86% by PCR analysis and 6.82%, 0, 47.06%, 34.69% by Southern blot analysis . The transgenic mice showed green fluorescence in fluorescence imager and fluorescent microscope under EGFP excitation light （488 nm）. Through comparing efficiency of transgenic mice in different spermiogenesis stages, it could provided an important theoretic foundation for efficient production of transgenic animals by testis mediated gene transfer.