受精卵原核显微穿刺导入DNA制备转基因兔

Introducing DNA into Zygotes through Nuclear Micro-Pricking to Produce Transgenic Rabbit

目的:受精卵原核显微DNA注射是转基因动物制备的最常用、最可靠的方法.研究技术上更为简易的受精卵原核显微穿刺导入DNA制备转基因兔的可行性.方法:重组转基因载体pCA-CHA,采用细胞原核显微穿刺将DNA导入兔受精卵制备转基因兔,用PCR和Southern Blot方法鉴定转基因兔.结果:转基因处理后获得仔兔92只,以PCR检测结果计算,转基因总效率为1.63%～2.40%,整合率为11.54%～16.13%;以Southern Blot检测结果计算,转基因总效率为0.27%,整合率为1.92%.结论:受精卵原核显微穿刺导入DNA与受精卵原核显微DNA注射制备转基因兔的结果相当.

Objective： Nuclear DNA micro-injection of zygotes is most common and most reliable method for transgenic mammalian production. We have successfully produced transgenic rabbits by nuclear micro- pricking to introduce DNA into zygotes （ a more simple and easy possessed method）. Methods. Expression vector pCA-CHA was constmcted and used for producing transgenic rabbits by nuclear DNA micro-pricking of zygote. Results： PCR and Southern blotting were applied for identification of transgenic rabbits. The total efficiency of the gene transfer was between 0.96% and 1.63% with imegration rates of 6.45% and 11.54% by PCR analysis. The total efficiency of the gene transfer was 0.27% with an integration rate of 1.92% determined by Southern blotting. Conclusion. These results are compatible with ones obtained by nuclear DNA micro- injection of rabbit zygotes.