Robson K J, Chandra T, MacGillivray R T, Woo S L
Proc Natl Acad Sci U S A. 1982 Aug;79(15):4701-5. doi: 10.1073/pnas.79.15.4701.
The mRNA for phenylalanine hydroxylase (phenylalanine 4-monooxygenase, EC 1.14.16.1) has been purified from total rat liver mRNAs, of which it constitutes less than 0.25%, to greater than 10% purity in a single step by specific polysome immunoprecipitation. The purified mRNA was used for synthesis and cloning of its cDNA. Recombinant colonies containing phenylalanine hydroxylase DNA sequences were identified by differential hybridization, hybrid-selected translation, and blot hybridization analysis. The rat cDNA clone was capable of hybridizing with human phenylalanine hydroxylase mRNA, which will permit the isolation of the corresponding human gene for analysis of phenylketonuria, a hereditary disorder in phenylalanine metabolism that causes permanent mental retardation in humans.
苯丙氨酸羟化酶（苯丙氨酸4-单加氧酶，EC 1.14.16.1）的信使核糖核酸（mRNA）已从大鼠肝脏总mRNA中纯化出来，在总mRNA中其含量不到0.25%，通过特异性多核糖体免疫沉淀一步法可将其纯度提高到10%以上。纯化后的mRNA用于其互补脱氧核糖核酸（cDNA）的合成和克隆。通过差异杂交、杂交选择翻译和印迹杂交分析，鉴定出含有苯丙氨酸羟化酶DNA序列的重组菌落。大鼠cDNA克隆能够与人苯丙氨酸羟化酶mRNA杂交，这将有助于分离相应的人类基因，用于分析苯丙酮尿症，这是一种苯丙氨酸代谢的遗传性疾病，可导致人类永久性智力迟钝。
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