Inoue S, Takahashi K, Ohta M
Tsukuba Research Institute, Banyu Pharmaceutical Co., Ltd., Tsukuba, Japan.
Genomics. 1999 Apr 1;57(1):169-72. doi: 10.1006/geno.1999.5750.
A method was developed for effective isolation of trinucleotide repeats from genomic DNA. This method is based on the DNA polymerase reaction, which is restricted with only two or three nucleotide substrates and primed by biotinylated oligonucleotide probes. Sequences are then isolated by a streptavidin biotin-trapping method. More than 80% of the clones from each library contained more than eight trinucleotide repeats. Sequence analysis showed that the characteristic dinucleotide flanking sequences usually confronting various trinucleotide repeats are not found in the vicinity of CAG repeats, suggesting that CAG repeats may have been generated through a mechanism different from that of other trinucleotide repeats.
开发了一种从基因组DNA中有效分离三核苷酸重复序列的方法。该方法基于DNA聚合酶反应，该反应仅受两种或三种核苷酸底物的限制，并由生物素化的寡核苷酸探针引发。然后通过链霉亲和素生物素捕获法分离序列。每个文库中超过80%的克隆包含八个以上的三核苷酸重复序列。序列分析表明，在CAG重复序列附近未发现通常与各种三核苷酸重复序列相对的特征性二核苷酸侧翼序列，这表明CAG重复序列可能是通过与其他三核苷酸重复序列不同的机制产生的。
