Meyne J, Moyzis R K
Los Alamos National Laboratory, New Mexico 87545.
Genomics. 1989 May;4(4):472-8. doi: 10.1016/0888-7543(89)90270-x.
The pericentric region of human chromosome 17 was targeted for specific in situ hybridization of the alphoid DNA subfamily enriched on this chromosome. A recombinant DNA clone containing the entire higher order chromosome 17 alphoid repeat preferentially hybridized to the pericentric region of chromosome 17, but frequently cross-hybridized to other chromosomes under normal stringency conditions. Chromosomal specificity, after in situ hybridization to metaphase spreads and interphase nuclei, was improved by using a subclone containing predominantly monomer 1 of the higher order repeat. Further improvement was achieved by synthesizing a 42-nucleotide oligomer of a divergent region of monomer 1. Southern blot analysis confirmed the improved specificity of the shorter probes. Reducing the potential of repetitive DNA probes to cross-hybridize increases the usefulness of the probes, especially when they are used for localizing individual chromosomes in interphase nuclei.
人类17号染色体的着丝粒周围区域是富含该染色体的α卫星DNA亚家族特异性原位杂交的目标区域。一个包含整个17号染色体高阶α卫星重复序列的重组DNA克隆优先与17号染色体的着丝粒周围区域杂交，但在正常严谨条件下经常与其他染色体发生交叉杂交。通过使用主要包含高阶重复序列单体1的亚克隆，在与中期染色体铺展和间期核进行原位杂交后，染色体特异性得到了提高。通过合成单体1不同区域的42个核苷酸的寡聚物，进一步提高了特异性。Southern印迹分析证实了较短探针的特异性提高。降低重复DNA探针交叉杂交的可能性增加了探针的实用性，特别是当它们用于在间期核中定位单个染色体时。
