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论文摘要

PiggyBac 转座子在肝脏中的活性研究

the activity of PiggyBac transposon in liver

作者:刘华华(四川大学生命科学学院功能基因组实验室);缪辉(四川大学生命科学学院功能基因组实验室);葛梦芸(四川大学生命科学学院功能基因组实验室);吴传芳(四川大学生命科学学院功能基因组实验室)

Author:LIU Hua-Hua(Department of Functional Genome, College of Life Science, Sichuan University);MIAO Hui(Department of Functional Genome, College of Life Science, Sichuan University);GE Meng-Yun(Department of Functional Genome, College of Life Science, Sichuan University);WU Chuan-Fang(Department of Functional Genome, College of Life Science, Sichuan University)

收稿日期:2015-05-08          年卷(期)页码:2016,53(5):1130-1134

期刊名称:四川大学学报: 自然科学版

Journal Name:Journal of Sichuan University (Natural Science Edition)

关键字:PiggyBac(PB)转座子;高压尾静脉注射;Splinkerette PCR;插入突变

Key words:PiggyBac(PB) transposon; Hydrodynamic injection; Splinkerette PCR; Insertional mutagenesis

基金项目:国家自然科学基金

中文摘要

PiggyBac(PB)转座子已经成为分析哺乳动物基因功能的有效工具,既可以作为转基因工具研究已知基因的功能,又可以作为插入突变的诱变剂研究未知基因的功能.之前关于PB转座子在体内的活性研究较少,且多研究其转基因功能;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系统可以作为有效地基因诱变工具来研究基因功能.

英文摘要

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.

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