IDENTIFICATION OF MUTATIONS LEADING TO THE LESCH-NYHAN SYNDROME BY AUTOMATED DIRECT DNA SEQUENCING OF INVITRO AMPLIFIED CDNA
IDENTIFICATION OF MUTATIONS LEADING TO THE LESCH-NYHAN SYNDROME BY AUTOMATED DIRECT DNA SEQUENCING OF INVITRO AMPLIFIED CDNA
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DOI：
10.1073/pnas.86.6.1919
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发表时间：
1989-03-01
影响因子：
11.1
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）综合征是一种严重的X染色体相关疾病，由嘌呤回收酶次黄嘌呤磷酸核糖基转移酶（HPRT）缺乏引起。导致疾病的突变是异质的，并且经常作为新生事件出现。我们通过对体外扩增的HPRT cDNA进行直接DNA测序，鉴定了15例独立产生的HPRT缺乏症的核苷酸改变。我们还证明了直接DNA序列分析可以自动化，进一步简化了该位点新突变的检测。突变包括DNA碱基替换、小DNA缺失、单个DNA碱基插入和RNA剪接错误。应用这些程序可以通过直接检测受LN影响的单个家庭中的突变等位基因来进行DNA诊断和携带者鉴定。
The Lesch-Nyhan (LN) syndrome is a severe X chromosome-linked disease that results from a deficiency of the purine salvage enzyme hypoxanthine phosphoribosyltransferase (HPRT). The mutations leading to the disease are heterogeneous and frequently arise as de novo events. We have identified nucleotide alterations in 15 independently arising HPRT-deficiency cases by direct DNA sequencing of in vitro amplified HPRT cDNA. We also demonstrate that the direct DNA sequence analysis can be automated, further simplifying the detection of new mutations at this locus. The mutations include DNA base substitutions, small DNA deletions, a single DNA base insertion, and errors in RNA splicing. The application of these procedures allows DNA diagnosis and carrier identification by the direct detection of the mtuant alleles within individual families affected by LN.