不同睾丸注射方法对转基因效率的影响  

Effects of testis injection methods on transgenic mouse production

作　　者：李翔[1,2,3] 戴建军[2,3] 吴彩凤[2,3] 张树山[2,3] 张廷宇[2,3] 马恒东[1] 

机构地区：[1]四川农业大学动物科技学院,雅安625014 [2]上海市农业科学院畜牧兽医研究所,上海201106 [3]上海市农业遗传育种重点实验室动物遗传工程研究室,上海201106

出　　处：《上海农业学报》2014年第1期54-57,共4页Acta Agriculturae Shanghai

基　　金：国家转基因生物新品种培育重大专项(2009ZX08006-014B;2008ZX08006-005);上海市农业科学院青年基金(QJ2012-07);上海市农委成果转化项目(123919N0700)资助

摘　　要：将pEGFP-C1质粒与脂质体混合后注入小鼠睾丸内,30d后观察睾丸形态和组织变化;检测精子和配种后仔鼠转基因阳性率,探讨小鼠睾丸网、曲精细管和睾丸间质3种注射方法对转基因小鼠生产效率的影响。结果显示:3种不同注射方法对小鼠睾丸造成损伤由大到小依次为睾丸网注射、曲精细管注射和睾丸间质注射。荧光检测小鼠精子,睾丸网和曲精细管注射法阳性率分别为(35.13±1.72)%和(15.13±1.45)%,睾丸间质注射法没有观察到阳性精子。PCR检测仔鼠,三者的转基因效率分别为33.3%、12.5%和0。The plasmid DNA（pEGFP-C1）mixed with liposome was injected into mouse testes,30 days later the morphological and histological changes of testes were observed, and the rates of both positive sperms and positive transgenic mice after joining were determined so as to compare the effects of three testis injection methods （rete testis injection, seminiferous tubules injection and testicular interstitial injection）on the productive efficiency of transgenic mice. The results showed that the descending order of testis damages by the above 3 methods was rete testis injection, seminiferous tubules injection and testicular interstitial injection. The fluorescence detection of mouse perms indicated that the positive sperm rates of rete testis injection, seminiferous tubes injection and testicular interstitial injection were（ 35.13 ± 1. 72）%, （15.13 ± 1.45）% and 0 respectively. The PCR detection of the progeny indicated that the positive transgenic mice rates by the 3 methods were 33.3%,12.5% and 0 respectively.

关 键 词：小鼠 转基因 睾丸注射 睾丸网注射 

分 类 号：S814.8[农业科学—畜牧学]