CYTOGENETIC ANALYSIS USING QUANTITATIVE, HIGH-SENSITIVITY, FLUORESCENCE HYBRIDIZATION
CYTOGENETIC ANALYSIS USING QUANTITATIVE, HIGH-SENSITIVITY, FLUORESCENCE HYBRIDIZATION
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DOI：
10.1073/pnas.83.9.2934
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发表时间：
1986-05-01
影响因子：
11.1
通讯作者：
GRAY, JW
中科院分区：
文献类型：
--
作者：
PINKEL, D;STRAUME, T;GRAY, JW
本文介绍了荧光原位杂交技术在染色体分类和染色体畸变检测中的应用。生物素标记的DNA与靶染色体杂交，随后通过用荧光素标记的亲和素和生物素化的抗亲和素抗体连续处理而呈现荧光。以人基因组DNA为探针，在人-仓鼠杂交细胞系中，人染色体在中期铺展和间期核中被强烈而均匀地染色。种间易位在中期很容易检测到。来自细胞核和染色体的人类特异性荧光强度与靶人类DNA的量成比例。用Y染色体特异的0.8-DNA酶探针对人Y染色体中期和间期核进行荧光染色。男性的细胞比女性的细胞亮40倍。两个Y染色体结构域可见于大多数间期核的XYY巨噬细胞。用1.5-二硫键酶DNA探针对人中期染色体上的28 S核糖体RNA基因进行了清晰的染色。
This report describes the use of fluorescence in situ hybridization for chromosome classification and detection of chromosome aberrations. Biotin-labeled DNA was hybridized to target chromosomes and subsequently rendered fluorescent by successive treatments with fluorescein-labeled avidin and biotinylated anti-avidin antibody. Human chromosomes in human-hamster hybrid cell lines were intensely and uniformly stained in metaphase spreads and interphase nuclei when human genomic DNA was used as a probe. Interspecies translocations were detected easily at metaphase. The human-specific fluorescence intensity from cell nuclei and chromosomes was proportional to the amount of target human DNA. Human Y chromosomes were fluorescently stained in metaphase and interphase nuclei by using a 0.8-kilobase DNA probe specific for the Y chromosome. Cells from males were 40 times brighter than those from females. Both Y chromosomal domains were visible in most interphase nuclei of XYY amniocytes. Human 28S ribosomal RNA genes on metaphase chromosomes were distinctly stained by using a 1.5-kilobase DNA probe.