Maekawa M, Sudo K, Li S S, Kanno T
Department of Laboratory Medicine, Hamamatsu University School of Medicine, Japan.
Hum Genet. 1991 Nov;88(1):34-8. doi: 10.1007/BF00204925.
Genomic DNA prepared from LDH-A-deficient whole blood was amplified by the polymerase chain reaction technique using two primers specific for the active human LDH-A gene. The amplified fragment was examined by direct agarose gel electrophoresis, and a deletion of 20 base pairs (bp) in exon 6 of the LDH-A gene was found. The results permitted a clear distinction between the homozygous mutant, the heterozygous mutant, and wild-type genotypes. Moreover, HinfI digestion and direct sequencing of the amplified product confirmed the results from direct agarose gel electrophoresis. Four families, including 18 individuals, were shown to contain the same mutation, that is a 20-bp deletion in exon 6. All genotypes were consistent with their biochemical phenotypes as evaluated by the ratio of LDH-B to LDH-A subunits in erythrocytes. Thus, all four known affected families in Japan have been shown to carry the same mutant gene, which may have been derived from a single mutational event.
从缺乏乳酸脱氢酶A（LDH-A）的全血中提取基因组DNA，使用针对活性人LDH-A基因的两种引物，通过聚合酶链反应技术进行扩增。扩增片段通过直接琼脂糖凝胶电泳进行检测，发现LDH-A基因外显子6中缺失了20个碱基对（bp）。这些结果能够清晰地区分纯合突变体、杂合突变体和野生型基因型。此外，对扩增产物进行HinfI酶切和直接测序，证实了直接琼脂糖凝胶电泳的结果。四个家族，包括18名个体，被证明含有相同的突变，即外显子6中20-bp的缺失。所有基因型与其红细胞中LDH-B与LDH-A亚基的比例所评估的生化表型一致。因此，日本所有四个已知的受影响家族都被证明携带相同的突变基因，该基因可能源自单一的突变事件。
