MULTIPLEX DNA DELETION DETECTION AND EXON SEQUENCING OF THE HYPOXANTHINE PHOSPHORIBOSYLTRANSFERASE GENE IN LESCH-NYHAN FAMILIES
MULTIPLEX DNA DELETION DETECTION AND EXON SEQUENCING OF THE HYPOXANTHINE PHOSPHORIBOSYLTRANSFERASE GENE IN LESCH-NYHAN FAMILIES
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DOI：
10.1016/0888-7543(90)90545-6
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发表时间：
1990-06-01
期刊：
影响因子：
4.4
通讯作者：
CASKEY, CT
中科院分区：
文献类型：
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作者：
GIBBS, RA;NGUYEN, PN;CASKEY, CT
Lesch-Nyhan(LN)综合征是一种遗传性的人类神经系统疾病，由次黄嘌呤磷酸核糖转移酶(HPRT)基因失活突变引起。人类HPRT基因座完整DNA序列的阐明使我们能够构建多个寡核苷酸引物集，用于同时扩增所有9个HRPT外显子。多重聚合酶链式反应提供了一种简便的检测HPRT外显子缺失的方法，反应产物可以通过直接自动荧光DNA测序来识别基因的细微变化。从15例独立的LN患者中发现了HPRT基因的改变，并进行了10个LN家系研究。该测序方法使用固体载体，操作简单，灵敏度高，是诊断LN的首选方法。LN杂合子可以在不参考患病男性的情况下诊断。此外，这些程序将对体细胞诱变研究有用。
The Lesch-Nyhan (LN) syndrome is a genetically lethal human neurological disease that results from mutations that inactivate the hypoxanthine phosphoribosyltransferase (HPRT) gene. The elucidation of the complete DNA sequence of the human HPRT gene locus has enabled the construction of multiple oligonucleotide primer sets for the simultaneous in vitro anplification of all nine HRPT exons. The multiplex polymerase chaina reaction provides a facile assay for the detection of HPRT exon deletions and the reaction products can be analyzed by direct automated fluorescent DNA sequencing to identify subtle alterations in the gene. Alterations have been identified in the HPRT genes from 15 independent LN cases, and 10 LN family studies were performed. The sequencing method uses solid supports and is sufficiently simple and sensitive to be a favored approach for LN diagnosis. LN heterozygotes can be diagnosed without reference to the affected male. In addition, these procedures will be useful for somatic mutagenesis studies.