Eubanks J H, Selleri L, Hart R, Rosette C, Evans G A
Molecular Genetics Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037.
Genomics. 1991 Nov;11(3):720-9. doi: 10.1016/0888-7543(91)90080-x.
A highly polymorphic repetitive sequence, D11S533, was isolated by oligonucleotide hybridization from an arrayed chromosome 11q-specific cosmid library. The DNA sequence of this element was determined and found to consist of a repetitive degenerate hexanucleotide sequence [T(Pu)T(Pu)T(Pu)]n extending over 438 bp. Southern blot analysis demonstrated that this element is relatively unique in the human genome. This sequence can be detected by amplification using the polymerase chain reaction (PCR) with oligonucleotide primers complementary to unique sequences flanking the repetitive element. This sequence displays a high degree of polymorphism, and analysis of 15 individuals demonstrated at least 10 alleles ranging in size from 300 to 900 bp. Fluorescence in situ hybridization was used to localize this sequence to 11q13 (FLpter 0.60 +/- 0.02). Pulsed-field gel electrophoresis and the isolation of yeast artificial chromosomes established the long-range physical map surrounding the locus. Because various alleles of this polymorphic sequence can be easily detected by PCR amplification, this probe has potential usefulness in genetic linkage mapping as well as identity testing.
通过寡核苷酸杂交从一个排列好的11号染色体q特异性黏粒文库中分离出一个高度多态的重复序列D11S533。测定了该元件的DNA序列，发现其由一个延伸438 bp的重复简并六核苷酸序列[T(Pu)T(Pu)T(Pu)]n组成。Southern印迹分析表明该元件在人类基因组中相对独特。使用与重复元件侧翼独特序列互补的寡核苷酸引物通过聚合酶链反应（PCR）扩增可以检测到该序列。该序列表现出高度多态性，对15个个体的分析显示至少有10个等位基因，大小在300至900 bp之间。荧光原位杂交用于将该序列定位到11q13（距pter 0.60 +/- 0.02）。脉冲场凝胶电泳和酵母人工染色体的分离建立了围绕该基因座的长距离物理图谱。由于该多态序列的各种等位基因可以通过PCR扩增轻松检测到，因此该探针在遗传连锁图谱绘制以及身份测试中具有潜在用途。
