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小鼠雌性生殖系统电穿孔与微泡增强超声波转EGFP基因的研究(简报)
引用本文:雷蕾,刘忠华,太田将,山田源.小鼠雌性生殖系统电穿孔与微泡增强超声波转EGFP基因的研究(简报)[J].分子细胞生物学报,2006,39(4).
作者姓名:雷蕾  刘忠华  太田将  山田源
作者单位:哈尔滨医科大学组织学与胚胎学教研室 哈尔滨150081(雷蕾),东北农业大学生命科学学院 哈尔滨150030(刘忠华),熊本大学生命资源研究支援中心 日本熊本8600811(太田将,山田源)
摘    要:非病毒载体转基因法,如注射裸DNA或脂质体转染,不产生细胞毒性,但除了肌肉组织外其他组织的转导效率均不高。电脉冲可使细胞膜产生临时的微孔允许一些分子通过,因此应用此方法可将药物或基因转入动物组织。电穿孔常用于培养的细胞转基因,理论上,低强度、长脉冲,或高强度、短脉冲有利于电转导,选择适合的参数是电转导的关键。本实验比较了不同电压和脉冲时间对小鼠卵巢在体转入绿色荧光蛋白基因的效果,确定了最适的电转导参数,为卵巢疾病的药物、基因治疗和研究卵泡发育中的基因调控提供了实验手段。超声波是临床常用的诊断方法,对人体无害…

关 键 词:电穿孔  超声波穿孔  绿色荧光蛋白  转导

IN VIVO TRANSDUCTION OF EGFP INTO FEMALE MOUSE REPRODUCTIVE SYSTEM BY ELECTROPORATION AND MICROBUBBLE-ENHANCED SONOPORATION
LEI Lei,LIU Zhong Hua,Ohta Sho,Yamada Gen.IN VIVO TRANSDUCTION OF EGFP INTO FEMALE MOUSE REPRODUCTIVE SYSTEM BY ELECTROPORATION AND MICROBUBBLE-ENHANCED SONOPORATION[J].Journal of Molecular Cell Biology,2006,39(4).
Authors:LEI Lei  LIU Zhong Hua  Ohta Sho  Yamada Gen
Abstract:In the present study,the efficiency of in vivo transduction of EGFP plasmid DNA into adult female mouse reproductive system by means of electroporation and microbubble-enhanced sonoporation are discussed. In the first experiment,EGFP plasmid DNA was injected into the ovary and following electroporation (square electric pulses were applied five times at 60V with a constant time of 20ms). Green fluorescence was detected 10h after transduction. In the second experiment, plasmid DNA was mixed with the microbubble at different ratio and injected into the uterus and the ovary separately. The tissues were exposed to different combined ultrasound parameters immediately after injection. We found that the suitable parameter was 2 w/cm2 intensity,20% duty cycle and 5min. Under this condition, green fluorescence was expressed in the uterus endometrium and ovarian cortex 12 h post-sonoporation and the expression lasted up to 5 days.
Keywords:Electroporation  Sonoporation  EGFP transduction
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