利用指甲DNA检测ALDH2基因的高通量诊断方法的建立

E. Nagayoshi, Y. Mizuta, Y. Takii
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引用次数: 0

摘要

利用指甲作为DNA材料来源,建立了一种有效筛选乙醛脱氢酶2 (ALDH2)基因的临床方法。通过对嗜碱性芽孢杆菌碱性蛋白酶基因的克隆和测序,在枯草芽孢杆菌中表达,获得了一种能溶解指甲角蛋白并灭活组织中共存的dna酶的高效蛋白酶。MIB029蛋白酶的氨基酸序列包含在其他四种枯草杆菌样蛋白酶中发现的共同区域。113名女大学生(平均年龄20.8±0.7岁;典型Glu纯合子ALDH2频率为0.66,杂合子(Glu487Lys)为0.32,非典型Lys纯合子为0.020。通过一份调查问卷,发现受试者以前没有收到有关其遗传背景与酒精饮料消费之间关系的信息。我们发现,通过收集指甲可以很容易地检测到酒精中毒的遗传单核苷酸多态性(SNP)背景,这对受试者或患者都很方便。
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Development of a High-Throughput Diagnosis Method for Detecting the ALDH2 Gene Using Fingernail DNA
A clinical method for effective genetic screening of the aldehyde dehydrogenase 2 (ALDH2) gene was developed, using the fingernail as a source of DNA material. A highly effective protease that could solubilize fingernail keratin and inactivate any DNase co-existing in the tissue was obtained by cloning and sequencing the gene for alkaline protease from Bacillus alcalophilus , followed by expression of the gene in Bacillus subtilis. The amino acid sequence of MIB029 protease contained common regions found in four other subtilisin-like proteases. In the fingernails of 113 female university students (average age 20.8 ± 0.7 years; body mass index, 20.4 ±1.6), ALDH2 frequency was 0.66 for the typical Glu homozygote, 0.32 for the heterozygote (Glu487Lys), and 0.020 for the atypical Lys homozygote. Through a questionnaire, it was found that the subjects had not previously received information regarding the relationship between their genetic background and consumption of alcoholic beverages. We found that the genetic single nucleotide polymorphism (SNP) background to alcoholism can be easily detected by collecting fingernails, which is convenient for subjects or patients.
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