Identification of inverted duplicated #15 chromosomes using bivariate flow cytometric analysis.
Identification of inverted duplicated #15 chromosomes using bivariate flow cytometric analysis.
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反向重复的识别
DOI：
10.1002/cyto.990060102
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
S. Latt
中科院分区：
文献类型：
--
作者：
M. Lalande;R. Schreck;R. Hoffman;S. Latt
双激光流式细胞仪IV细胞分选仪已被用来获得双变量流直方图的人中期染色体染色的DNA特异性染料，33258 Hoechst和色霉素A3。使用这种双重染色技术和流式细胞仪可以分辨大约20个不同的染色体荧光群体，流式细胞仪仅通过用专门设计的空气间隔消色差透镜代替标准聚焦透镜来进行修改。来自两个不同细胞系的中期染色体携带反向重复的#15常染色体，已进行双变量染色体分析。在这两种情况下，反向重复的#15染色体已经在双变量流直方图中被识别。这一鉴定得到了实验的支持，在实验中，双重染色的染色体用netropsin或偏端霉素A复染，导致结构异常的#15染色体的33258 Hoechst荧光强度相对增加，与其他染色体相比，如细胞学研究所预测的。使用市售仪器鉴定和分离小的异常常染色体的可能性应便于使用重组DNA技术构建文库，所述文库高度富集来自在染色体异常如缺失、易位和倒位重复的研究中重要的有限常染色体亚区的DNA序列。
A dual laser FACS IV cell sorter has been used to obtain bivariate flow histograms of human metaphase chromosomes stained with the DNA-specific dyes, 33258 Hoechst and chromomycin A3. Approximately twenty distinct chromosomal fluorescence populations can be resolved using this double staining technique and the flow cytometer which has been modified only by the substitution of a specially designed air-spaced achromat for the standard focusing lens. Metaphase chromosomes from two different cell lines bearing inverted duplicated #15 autosomes have been subjected to bivariate chromosome analysis. In both cases, the inverted duplicated #15 chromosomes have been identified in the bivariate flow histogram. This identification was supported by experiments in which doubly stained chromosomes were counterstained with either netropsin or distamycin A, resulting in a relative increase in the 33258 Hoechst fluorescence intensity of the structurally abnormal #15 chromosomes, compared with the other chromosomes, as predicted by cytological studies. The possibility of identifying and separating small abnormal autosomes using commercially available instrumentation should facilitate the use of recombinant DNA techniques for the construction of libraries which are highly enriched for DNA sequences from limited autosomal subregions important in the study of chromosomal abnormalities such as deletions, translocations and inversion duplications.