CAJ | 학술논문

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.
건립료일충전신이고효적제비전기인동물신방법.무수외과수술,장함유록색형광단백적중조질립직접다차다점반복주사도웅성ICR소서적고환내.궤주후,상술웅성동물여자연발정적자성교배,제비전기인동물.경PCR검측、DNA인적,실험결과표명:F1대소서전기인양성솔위41%.경DNA인적증명:외원기인이경정합도자대전기인동물적기인조내병능유전급후대.장F1대양성서여정상ICR서교배,산생F2대전기인서,F2대전기인양성솔37%.상술실험결과표명,건립적체내계통전기인방법간편、고효,괄용우대규모제비전기인동물,특별괄용우일사대형가축.
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 F1 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 of transgenic animals, especially, for producing domestic animals.