体内系统转基因新方法(英文)  

A Novel Method to Transfer Gene In vivo System

作　　者：何新[1,2] 齐冰[1,2] 刘桂生[1] 郁卫东[1] 陈清轩[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所 [2]中国科学院研究生院,北京100049

出　　处：《生物化学与生物物理进展》2006年第7期685-690,共6页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展计划(973)资助项目(G2000016107)~~

摘　　要：建立了一种全新而高效的制备转基因动物新方法.无需外科手术,将含有绿色荧光蛋白的重组质粒直接多次多点反复注射到雄性ICR小鼠的睾丸内.几周后,上述雄性动物与自然发情的雌性交配,制备转基因动物.经PCR检测、DNA印迹,实验结果表明:F1代小鼠转基因阳性率为41%.经DNA印迹证明:外源基因已经整合到子代转基因动物的基因组内并能遗传给后代.将F1代阳性鼠与正常ICR鼠交配,产生F2代转基因鼠,F2代转基因阳性率37%.上述实验结果表明,建立的体内系统转基因方法简便、高效,适用于大规模制备转基因动物,特别适用于一些大型家畜.A new and effective method to produce transgenic animals was established. Without a surgical incision, the recombinant plasmid containing green fluorescence protein （GFP） cDNA was repeatedly injected into male mouse testis at multi-sites. After few weeks of the final injection, the injected male was mated with normal oestrus female to produce transgenic mice. The presence of the GFP cDNA in F1 transgenic individuals were detected by polymerase chain reaction and Southern blot hybridization, which showed that the transgenic rate of mouse F） offspring was 41%. The transferred gene was integrated into the host genome and could be transmitted to its offspring. When the positive F1 individuals were mated with the wild type ICR mice, the F2 individuals had a transgenic rate of 37%. The results indicate that the high efficiency of gene transfer and the limited number of manipulations make the method suitable for creating a large number oftransgenic animals, especially, for producing domestic animals.

关 键 词：睾丸注射 基因转移 体内 转基因动物 

分 类 号：Q78[生物学—分子生物学] N94[自然科学总论—系统科学]