Fukao T, Yamaguchi S, Orii T, Osumi T, Hashimoto T
Department of Pediatrics, Gifu University School of Medicine, Japan.
Biochim Biophys Acta. 1992 Jul 7;1139(3):184-8. doi: 10.1016/0925-4439(92)90132-7.
3-Ketothiolase deficiency (3KTD) is the result of a deficiency in mitochondrial acetoacetyl-CoA thiolase (T2). The molecular basis of 3KTD was analyzed in a patient (GK10) and his family at the protein, cDNA and gene levels. Protein analyses showed that GK10's T2 protein was undetectable in fibroblasts even with the pulse-protein labeling method and that his parents were carriers of 3KTD. Complementary DNA analyses with PCR showed that T2 cDNA in the patient lacked the normal exon 11 sequence and that his parents were obligatory carriers of the DNA sequence which canceled exon 11. When the PCR-amplified genomic fragments around exon 11 were sequenced, an AG to AC mutation at the 3' splice site of intron 10 was detected. This mutation is presumed to be responsible for exon 11 skipping.
3-酮硫解酶缺乏症（3KTD）是线粒体乙酰乙酰辅酶A硫解酶（T2）缺乏的结果。在一名患者（GK10）及其家族中，从蛋白质、cDNA和基因水平分析了3KTD的分子基础。蛋白质分析表明，即使采用脉冲蛋白质标记法，在成纤维细胞中也检测不到GK10的T2蛋白，且其父母为3KTD携带者。PCR互补DNA分析显示，患者的T2 cDNA缺乏正常的外显子11序列，其父母为消除外显子11的DNA序列的 obligatory 携带者。对第11外显子周围的PCR扩增基因组片段进行测序时，在内含子10的3'剪接位点检测到AG到AC的突变。推测该突变是外显子11跳跃的原因。 （注：这里“obligatory carriers”似乎不太准确，可能是“杂合子携带者”之类的意思，但按要求未修改原文表述）
