中文摘要：

受精卵雄原核裸DNA注射是目前制备转基因小鼠的主要技术,而转基因表达成功率低是这种技术的主要缺点。piggyBac转座子系统已被报道用于制备转基因小鼠,但这一方法是否能够提高转基因的表达成功率尚不清楚。为此,我们利用毛色基因agouti为报告基因,采用piggyBac转座子系统以C57/BL6小鼠为背景进行转基因小鼠的制备。结果表明,将piggyBac转座酶cRNA和转基因载体进行受精卵雄原核共注射后,转基因阳性率为18.4%,转基因表达率为88.89%,显著高于单独进行转基因载体DNA受精卵雄原核注射法。同时,利用agouti基因作为报告基因,可根据毛色变化直接对表达阳性的转基因小鼠进行初步筛选,提高了筛选效率。

英文摘要：

Low transgene expression ratio in transgenic mice produced by male pronucleus injection of naked DNA into fertilized egg limits its application in various research purposes. In this paper, we used transposon piggyBac mediated transgenic method to prepare transgenic mice. The transgenic efficiency and transgene expression ratio were compared with that in traditional transgenic mouse preparation method. The agouti gene was chosen as transgene, driven by PGK promoter and cloned into the piggyBac transposon cassette of plasmid pB232. The transgenic positive rate was 18.4%. 8 of 9 transgenic mouse founders showed agouti fur color phenotype which was much higher in ratio than that of traditional method. The results also showed that agouti gene ectopic expression could lead to obesity which was consistent with previous study. In conclusion, our results indicated that comparing with the traditional naked DNA pronucleus microinjection, piggyBac-mediated transgenic mouse approach could significantly improve the expression of transgene in transgenic mice. Furthermore, using our report vector system, transgene expression positive transgenic mouse could be easily selected according to fur color.