Breneman J W, Swiger R R, Ramsey M J, Minkler J L, Eveleth J G, Langlois R A, Tucker J D
Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, CA 94551-9906.
Cytogenet Cell Genet. 1995;68(3-4):197-202. doi: 10.1159/000133913.
The recent development of mouse chromosome painting probes for fluorescence in situ hybridization has extended the use of this common laboratory mammal in cytogenetics. We now report the development of additional painting probes by degenerate-oligonucleotide-primed PCR on chromosomes from mouse lung fibroblast cultures, each homozygous for a single Robertsonian translocation chromosome. These probes are for Rb(1.2), Rb(1.3), Rb(4.6), and Rb(6.7). Probes were also made for the sex chromosomes by isolating shoulders from larger peaks (X) or small, clearly resolved peaks (Y) in the flow karyotype. Combinations of probes were used to paint four chromosomes simultaneously in a single color. Multicolor painting was achieved with a biotinylated Rb(1.2) probe and a digoxigenin-labeled Rb(2.8) probe. Each of the three different homologous pairs was uniquely colored by avidin-Texas Red, anti-digoxigenin-FITC, or both simultaneously. These results extend the usefulness of the mouse as a model for understanding adverse environmental exposures and genetic diseases in humans.
用于荧光原位杂交的小鼠染色体涂染探针的最新进展，拓展了这种常见实验哺乳动物在细胞遗传学中的应用。我们现在报告通过简并寡核苷酸引物PCR，从小鼠肺成纤维细胞培养物的染色体上开发额外的涂染探针，每个培养物对于单个罗伯逊易位染色体均为纯合子。这些探针针对Rb(1.2)、Rb(1.3)、Rb(4.6)和Rb(6.7)。还通过从流式核型中较大峰（X）或小的、清晰分辨的峰（Y）分离肩部，制备了性染色体的探针。探针组合用于在单一颜色中同时涂染四条染色体。用生物素化的Rb(1.2)探针和地高辛配体标记的Rb(2.8)探针实现了多色涂染。三个不同同源对中的每一对都通过抗生物素蛋白-德克萨斯红、抗地高辛配体-异硫氰酸荧光素或两者同时进行独特的染色。这些结果拓展了小鼠作为理解人类不良环境暴露和遗传疾病模型的用途。
