Hamaguchi T, Nakajima H, Noguchi T, Ono A, Kono N, Tarui S, Kuwajima M, Matsuzawa Y
Second Department of Internal Medicine, Osaka University Medical School, Suita, Japan.
Biochem Biophys Res Commun. 1994 Jul 15;202(1):444-9. doi: 10.1006/bbrc.1994.1948.
A genetic defect was investigated in a newly diagnosed Japanese case with muscle type phosphofructokinase (PFK-M) deficiency. Polymerase chain reaction (PCR) amplification of patient cDNA revealed an in-frame truncation of 165 bases. This was compatible to the complete deletion of exon 19. The rest of the sequence was identical to that of the normal PFK-M cDNA. Sequencing of PCR amplified genomic DNA of the patient revealed a point mutation from G to A at the 5' donor site of intron 19. This mutation resulted in the skipping of exon 19 in the patient mRNA. Homozygosity of this patient was confirmed by allele specific amplification of the genomic DNA. Donor mutations in intron 15 and intron 5 associated with different splicing errors were previously reported to cause this disease. Thus, the human PFK-M gene mutations are heterogeneous, however, the donor mutations and splicing errors would represent one of the frequent causes of this disease.
对一名新诊断出的日本肌肉型磷酸果糖激酶（PFK-M）缺乏症患者的基因缺陷进行了研究。对患者的cDNA进行聚合酶链反应（PCR）扩增，结果显示有165个碱基的框内截短。这与外显子19的完全缺失相符。其余序列与正常PFK-M cDNA的序列相同。对患者PCR扩增的基因组DNA进行测序，结果显示内含子19的5'供体位点发生了从G到A的点突变。该突变导致患者mRNA中外显子19的跳跃。通过基因组DNA的等位基因特异性扩增证实了该患者的纯合性。先前有报道称，内含子15和内含子5中的供体突变与不同的剪接错误相关，可导致这种疾病。因此，人类PFK-M基因突变具有异质性，然而，供体突变和剪接错误可能是这种疾病的常见病因之一。
