应用荧光原位杂交检测人凝血因子Ⅸ在转基因小鼠染色体上的整合  

Detection of the Integration of Human FⅨ(hFⅨ)on Chromosomes of Transgenic Mice by Fluorescence in situ Hybridization

作　　者：肖艳萍[1] 奚鹰[1] 黄文英[1] 黄英[1] 

机构地区：[1]上海市儿童医院,上海医学遗传研究所上海200040

出　　处：《遗传》2002年第3期232-236,共5页Hereditas（Beijing)

基　　金：国家"8 6 3"高技术项目 (编号 2 0 0 1AA2 130 1);上海市现代生物与新药产业发展基金 ( 9943190 4)资助

摘　　要：应用荧光原位杂交 (FISH)技术检测两个转基因小鼠家系从F1到F4 代的整合情况。阳性转基因小鼠 98%～ 10 0 %的中期分裂相 ,85 %～ 94%的间期核出现杂交信号 ;阴性对照小鼠 10 0 %的中期分裂相、95 %～ 96 %的间期核未出现杂交信号。结果表明 ,该FISH实验条件能对转基因整合位点进行高效特异检测。本文分析的两家系转基因小鼠均为单位点整合 ,但整合位点不同。各家系内F1到F4 代的转基因小鼠均可检出整合染色体 ,且整合位点相同 ,表明外源基因稳定整合并遗传给后代。Fluorescence in situ hybridization (FISH) was used to detect the integration of hFⅨ on chromosomes of transgenic mice from F 1 to F 4 generation in two strains.For transgenic mice,98%～100% of metaphases and 85%～94% of interphases showed hybridization signal.For negative control mice,100% of metaphases and 95%～96% of interphases showed no hybridization signal.The results demonstrated that FISH developed to detect the integration sites of hFⅨ was high efficient and specific.The integration sites of the transgenic mice analyzed were both single but different between the two strains.The integration chromosomes can be found in the transgenic mice from F 1 to F 4 generation and the integration sites were the same as each of the strains,which indicated that the transgene was stably integrated and transmitted to offspring.

关 键 词：荧光原位杂交 检测 人凝血因子Ⅸ 转基因小鼠 染色体 整合位点 

分 类 号：Q78[生物学—分子生物学]