PiggyBac 转座子在肝脏中的活性研究

本实验运用高压尾静脉注射的方法将PB转座系统导入小鼠肝脏,研究其在肝脏中的转座活性,系统中PB转座子携带表达红色荧光蛋白的基因以指示转座的发生.注射10个月后取出小鼠肝脏,体视荧光显微镜观察肝脏是否有红色荧光,并通过基因组PCR、splinkerette PCR分析PB转座子在肝脏中的插入情况.实验结果表明,PB转座子在肝脏内发生高效转座,检测的68个PB插入位点中有34个位于基因序列,使得PB系统可以作为有效的基因诱变工具来研究基因功能.

the activity of PiggyBac transposon in liver

PiggyBac(PB) transposon has become an effective tool for studying gene function, for it can be used for transgene and insertional mutagesis to explore gene function. Previous studies on the activity of PB transposon are mainly in vitro, and they primarily focused on transgenesis. In this study, PB binary cotransfection system was direct-ly delivered into liver via hydrodynamic injection using RFP (Red Fluorescence Protein) as the indicator of PB transposition. Tests were run 10 months later to detect whether the liver emitted red fluorescent. Then, genomic PCR and splinkerette PCR were performed to analyze the insertion sites of PB transposon. The results showed that PB transposon has an efficient transposition in liver as 34/68 insertion sites were located in gene sequence.