Gene mapping on human metaphase chromosomes by in situ hybridization with 3H, 35S, and 32P labeled probes and transmission electron microscopy.
Gene mapping on human metaphase chromosomes by in situ hybridization with 3H, 35S, and 32P labeled probes and transmission electron microscopy.
复制标题
通过使用 3H、35S 和 32P 标记探针和透射电子显微镜进行原位杂交，对人类中期染色体进行基因定位。
DOI：
10.1007/bf00327588
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发表时间：
1986
期刊：
影响因子：
1.6
通讯作者：
Davidson,N
中科院分区：
文献类型：
--
作者：
Li,CB;Wu,M;Margitich,IS;Davidson,N
提出了一种基于原位杂交、放射自显影和透射电子显微镜的人类中期染色体基因定位方法。描述了串联重复 rDNA 基因和两个核酸探针 N-mycand 探针 3（Kanda 等人，1983）的成功作图，这些探针在神经母细胞瘤细胞系 IMR-32 的均匀染色区域（HSR）中扩增。通过使用足够薄的AgBr乳液，可以用3H、35S或32P放射性标记的探针获得可观察的颗粒和良好的分辨率，但前者给出最好的结果。我们观察到，N-mycand 和探针 3 这两个探针均沿 HSR 没有均匀的空间分布，并且两个探针的分布彼此不同。这些观察结果支持了先前的研究，这些研究表明 HSR 的形成是一个比均匀扩增单个 DNA 片段以形成 n 倍完美串联重复序列更为复杂的过程。本研究表明，电子显微镜方法有助于扩展光学显微镜研究结果，解决需要更高分辨率绘图的问题。
A method based on in situ hybridization, autoradiography and transmission electron microscopy for mapping genes on human metaphase chromosomes is presented. Successful mapping of the tandemly repeated rDNA genes and of two nucleic acid probes, N-mycand probe 3 (Kanda et al. 1983), that are amplified in a homogeneously staining region (HSR) of the neuroblastoma cell line, IMR-32 is described. By using sufficiently thin AgBr emulsions, it is possible to obtain observable grains and good resolution with probes radiolabeled with3H,35S, or32P, but the former gives the best results. We observe that neither of the two probes, N-mycand probe 3, has a uniform spatial distribution along the HSR and that the distributions of the two probes differ from each other. These observations support previous studies which indicated that the formation of an HSR is a more complex process than uniform amplification of a single DNA segment to form an n-fold set of perfect tandem repeats. The present study shows that the electron microscopic method is useful for extending the results of light microscopic studies for problems where higher resolution mapping is needed.