Brown S, Wiebel F J, Gelboin H V, Minna J D
Proc Natl Acad Sci U S A. 1976 Dec;73(12):4628-32. doi: 10.1073/pnas.73.12.4628.
Hybrid clones segregating human chromosomes were prepared by fusing mouse RAG cells to fresh human bone marrow cells and tested for the mixed-function oxygenase [flavoprotein-linked monooxygenase; RH, reduced-flavoprotein:oxygen oxidoreductase (RH-hydroxylating); EC 1.14.14.1] arylhydrocarbon hydrocarbon hydroxylase. Neither constitutive nor induced aryl hydrocarbon hydroxylase activity was detected in parental RAG cells. Induced aryl hydrocarbon hydroxylase was expressed in 4 out of 12 primary and 12 out of 19 secondary hybrid clones examined. Constitutive hydroxylase activity was detectable in 9 of the 15 inducible clones. All of the hybrid clones that exhibited constitutive hydroxylase activity were also inducible. There was a positive correlation between constitutive and induced hydroxylase activities although the absolute levels of the enzyme showed a wide range between different clones. Isozyme analysis performed on 12 primary and 19 secondary hybrid clones showed that aryl hydrocarbon hydroxylase activity was concordant with the expression of the human isozymes malate dehydrogenase (EC 1.1.1.37) and isocitrate dehydrogenase (EC 1.1.1.42), previously assigned to human chromosome 2. Isozyme markers for 19 other human chromosomes segregated independently from aryl hydrocarbon hydroxylase activity. The results suggest that the gene(s) required for aryl hydrocarbon hydroxylase activity are located on human chromosome 2.
通过将小鼠RAG细胞与新鲜人骨髓细胞融合制备了分离人染色体的杂种克隆，并对其进行了混合功能加氧酶[黄素蛋白连接的单加氧酶；RH，还原型黄素蛋白：氧氧化还原酶（RH-羟化）；EC 1.14.14.1]芳烃羟化酶的检测。在亲本RAG细胞中未检测到组成型或诱导型芳烃羟化酶活性。在所检测的12个初级杂种克隆中有4个以及19个次级杂种克隆中有12个表达了诱导型芳烃羟化酶。在15个可诱导克隆中的9个中可检测到组成型羟化酶活性。所有表现出组成型羟化酶活性的杂种克隆也都是可诱导的。尽管不同克隆中该酶的绝对水平差异很大，但组成型和诱导型羟化酶活性之间存在正相关。对12个初级杂种克隆和19个次级杂种克隆进行的同工酶分析表明，芳烃羟化酶活性与先前定位于人染色体2的人同工酶苹果酸脱氢酶（EC 1.1.1.37）和异柠檬酸脱氢酶（EC 1.1.1.42）的表达一致。其他19条人染色体的同工酶标记与芳烃羟化酶活性独立分离。结果表明，芳烃羟化酶活性所需的基因位于人染色体2上。
