【摘要】 由于传统显微注射法具有操作繁琐、技术要求高和外源基因导入率低等局限性,本试验对常规显微注射法进行改进,旨在建立一套比较完善的水平显微注射方法。采用水平微注射系统,将外源DNA片段导入胚胎的雄原核中,获得子代鼠,分娩后3周提取鼠尾基因组DNA,PCR法筛选转基因阳性鼠。本试验采用Puc/BLG IN S和Chym os in基因,共使用710枚鼠原核胚进行研究,胚胎经注射后移植了25只受体,产下仔鼠77只;妊娠率分别为33.3%(5/15)和50%(5/10),胚胎成活率分别为9.47%(41/433)和l3.00%(36/277),阳性率分别为4.88%(2/41)和8.33%(3/36)。将阳性雌鼠配种妊娠后,对外源基因整合情况进行检测,结果表明初步获得了转基因阳性小鼠。更多还原

【Abstract】 The purpose of this study is to produce transgenic mice integrated with foreign genes.Foreign genes were introduced into the zygote of mice by horizontal microinjection.Puc/BLGINS and Chymosin genes were seperately injected into male pronucleus of 710 zygotes and injected eggs were implanted into 25 foster mothers,which bore 77 offsprings.Genomic DNA was extracted from the tail of mice obtained from the founder mice for PCR analysis.The pregnant rates were seperately 33.3%(5/15) and 50%(5/10).The embryo rates were seperately 9.47%(41/433) and 13.00%(36/277).The positive rates of transgenic mice was seperately 4.88%(2/41) and 8.33%(3/36).The model of transgenic mice was established primarily.更多还原