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用合成串联重复探针检测人类小卫星基因座:与克隆DNA指纹图谱探针的直接比较

Isolation of human minisatellite loci detected by synthetic tandem repeat probes: direct comparison with cloned DNA fingerprinting probes.

作者信息

Armour J A, Vergnaud G, Crosier M, Jeffreys A J

机构信息

Department of Genetics, University of Leicester, UK.

出版信息

Hum Mol Genet. 1992 Aug;1(5):319-23. doi: 10.1093/hmg/1.5.319.

Abstract

As a direct comparison with cloned 'DNA fingerprinting' probes, we present the results of screening an ordered array Charomid library for hypervariable human loci using synthetic tandem repeat (STR) probes. By recording the coordinates of positive hybridization signals, the subset of clones within the library detected by each STR probe can be defined, and directly compared with the set of clones detected by naturally occurring (cloned) DNA fingerprinting probes. The STR probes vary in the efficiency of detection of polymorphic minisatellite loci; among the more efficient probes, there is a strong overlap with the sets of clones detected by the DNA fingerprinting probes. Four new polymorphic loci were detected by one or more of the STR probes but not by any of the naturally occurring repeats. Sequence comparisons with the probe(s) used to detect the locus suggest that a relatively poor match, for example 10 out of 14 bases in a limited region of each repeat, is sufficient for the positive detection of tandem repeats in a clone in this type of library screening by hybridization. These results not only provide a detailed evaluation of the usefulness of STR probes in the isolation of highly variable loci, but also suggest strategies for the use of these multi-locus probes in screening libraries for clones from hypervariable loci.

摘要

作为与克隆的“DNA指纹”探针的直接比较,我们展示了使用合成串联重复序列(STR)探针筛选有序阵列鲱鱼精DNA文库以寻找人类高变位点的结果。通过记录阳性杂交信号的坐标,可以确定每个STR探针在文库中检测到的克隆子集,并直接与天然存在的(克隆的)DNA指纹探针检测到的克隆集进行比较。STR探针在检测多态性小卫星位点的效率上有所不同;在效率较高的探针中,与DNA指纹探针检测到的克隆集有很强的重叠。一种或多种STR探针检测到了四个新的多态性位点,但任何天然存在的重复序列都未检测到。与用于检测该位点的探针进行序列比较表明,在这种类型的文库杂交筛选中,相对较差的匹配(例如每个重复序列有限区域内14个碱基中的10个)就足以在克隆中阳性检测串联重复序列。这些结果不仅详细评估了STR探针在分离高变位点方面的有用性,还提出了在筛选文库以寻找来自高变位点的克隆时使用这些多位点探针的策略。

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