Abadie V, Jaruzelska J, Lyonnet S, Millasseau P, Berthelon M, Rey F, Munnich A, Rey J
Unité de Recherches sur les Handicaps Génétiques de l'Enfant, INSERM U12, Paris, France.
Hum Mol Genet. 1993 Jan;2(1):31-4. doi: 10.1093/hmg/2.1.31.
Taking advantage of the 'illegitimate' transcription of the phenylalanine hydroxylase (PAH) gene, we have been able to analyse the PAH cDNA sequence of hyperphenylalaninemic children in circulating lymphocytes. Using this approach, we have also identified 3 novel mutations in cDNA from liver and lymphocytes of two patients. One mutation, detected by the abnormal pattern of migration of an amplified fragment, is a C to T transition in the splice acceptor site of intron 10, which resulted in the skipping of exon 11 with the premature termination of RNA translation downstream from exon 12 (-3 IVS10). The other two mutations are missense mutations in exons 10 and 11 (respectively, L333F and E390G). The present study supports the view that circulating lymphocytes give easy access to PAH gene transcripts whose nucleotide sequence is identical to that reported in liver and therefore represent a useful tool for molecular genetic studies in phenylketonuria.
利用苯丙氨酸羟化酶（PAH）基因的“非法”转录，我们得以分析高苯丙氨酸血症儿童循环淋巴细胞中的PAH cDNA序列。通过这种方法，我们还在两名患者的肝脏和淋巴细胞cDNA中鉴定出3个新突变。其中一个突变是通过扩增片段异常迁移模式检测到的，为内含子10剪接受体位点的C到T转换，导致外显子11跳跃，外显子12下游的RNA翻译提前终止（-3 IVS10）。另外两个突变是外显子10和11中的错义突变（分别为L333F和E390G）。本研究支持这样一种观点，即循环淋巴细胞便于获取PAH基因转录本，其核苷酸序列与肝脏中报道的相同，因此是苯丙酮尿症分子遗传学研究的有用工具。
