| 以质粒为载体的小片段DNA做荧光原位杂交进行基因定位及检测 |
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| 引用本文： | 郑淑芳 乔惠珍. 以质粒为载体的小片段DNA做荧光原位杂交进行基因定位及检测[J]. 中华肿瘤杂志, 1999, 21(5): 339-341 |
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| 作者姓名： | 郑淑芳 乔惠珍 |
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| 作者单位： | 北京医科大学临床肿瘤学院北京市肿瘤防治研究所遗传室(郑淑芳!100034，路桂荣!100034，冯莉雅!100034，宁涛!100034)，呼和浩特市内蒙古医学院第一附属医院妇产科(乔惠珍)，北京医科大(柯杨!100034) |
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| 基金项目： | 国家自然科学基金，863 国家重点攻关项目 |
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| 摘 要： | 目的 建立以质粒为载体小片段ＤＮＡ的荧光原位杂交（ＦＩＳＨ）基因定位及检测方法。方法 采用切口平移法以载有目的基因的质粒直接标记生物素制成探针，用ＦＩＳＨ法于外周血淋巴细胞中期分裂相定位新基因１Ａ６在卵巢癌和胃癌细胞株上检测癌基因ｃ－ｍｅｔ，ｃ－ｅｒｂＢ２，１Ａ６和抑癌基因ＡＰＣ，Ｒｂ。结果 新基因１Ａ６初步定位于Ｃ组某一染色体长臂远端，上述癌基因，抑癌基因在卵巢癌和胃癌细胞株中的获得清晰的杂交信
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| 关 键 词： | 质粒 DNA 荧光原位杂交 基因定位 基因检测 |
Gene detection and localization by FISH with short DNA fragment cloned in plasmid |
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| ZHENG Shufang,QIAO Huizhen,LU Guirong,et al.. Gene detection and localization by FISH with short DNA fragment cloned in plasmid[J]. Chinese Journal of Oncology, 1999, 21(5): 339-341 |
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| Authors: | ZHENG Shufang QIAO Huizhen LU Guirong et al. |
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| Affiliation: | Beijing Institute for Cancer Research, School of Oncology, Beijing Medical University, Beijing 100034. |
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| Abstract: | Objective To establish a method of detecting and localizing newly cloned gene by FISH using short DNA fragment. Methods Bio 14 dATP was incorporated into the target gene cloned in plasmid by nick translation followed by conventional FISH with some modification. By Means of this method, c met, c erbB2, 1A6, APC and Rb genes were examined in ovarian and gastric cancer cell lines. Results 1A6, a newly isolated gene was located at the long arm of one of the C group chromosomes. Oncogene c met, c erbB2 and trmor suppressor gene APC, Rb were shown to produce clear cut hybridization signals in ovarian and gastric cancer cells. Conclusion Short DNA fragment cloned in plasmid is useful for studies of gene localization, gene deletion and amplification by FISH. |
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| Keywords: | Plasmid DNA In situ hybridization fluorescence Gene localization Gene detection |
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