D'Ancona G G, Wurm J, Croce C M
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4526-9. doi: 10.1073/pnas.76.9.4526.
We have studied somatic cell hybrids between thymidine kinase (EC 2.7.1.75) deficient mouse cells and human diploid fibroblasts for the expression of human acid alpha-glucosidase (EC 3.2.1.20). A deficiency in this enzyme is associated with the type II glycogenosis or Pompe disease. All 30 somatic cell hybrids selected in hypoxanthine/aminopterin/thymidine medium expressed human acid alpha-glucosidase and galactokinase (EC 2.7.1.6) and retained human chromosome 17; counterselection of the same hybrids in medium containing 5-bromodeoxyuridine resulted in the growth of hybrids that concordantly lost the expression of human acid alpha-glucosidase and galactokinase as well as human chromosome 17. Hybrids between thymidine kinase-deficient mouse cells and fibroblasts from a patient with Pompe disease that contained human chromosome 17 were found not to express human acid alpha-glucosidase. Because we have already shown that hybrids between mouse peritoneal macrophages and GM54VA simian virus 40-transformed human cells selectively retain human chromosome 17 and lose all other human chromosomes, we tested 13 independent mouse macrophage x GM54VA hybrid clones, including two that retained human chromosome 17 and no other human chromosomes, for the expression of human acid alpha-glucosidase and galactokinase. All 13 hybrid clones were found to express these human enzymes. Thus, we conclude that the gene coding for human acid alpha-glucosidase is located on human chromosome 17.
我们研究了胸苷激酶(EC 2.7.1.75)缺陷的小鼠细胞与人二倍体成纤维细胞之间的体细胞杂种,以检测人酸性α-葡萄糖苷酶(EC 3.2.1.20)的表达情况。该酶的缺乏与II型糖原贮积病或庞贝病相关。在次黄嘌呤/氨基蝶呤/胸腺嘧啶核苷培养基中筛选出的所有30个体细胞杂种均表达人酸性α-葡萄糖苷酶和半乳糖激酶(EC 2.7.1.6),并保留了人类17号染色体;在含有5-溴脱氧尿苷的培养基中对相同杂种进行反选择,结果导致杂种生长,这些杂种同时丧失了人酸性α-葡萄糖苷酶和半乳糖激酶的表达以及人类17号染色体。发现胸苷激酶缺陷的小鼠细胞与来自一名庞贝病患者的成纤维细胞之间含有人类17号染色体的杂种不表达人酸性α-葡萄糖苷酶。由于我们已经表明小鼠腹腔巨噬细胞与GM54VA猿猴病毒40转化的人类细胞之间的杂种选择性地保留人类17号染色体并丢失所有其他人类染色体,因此我们检测了13个独立的小鼠巨噬细胞×GM54VA杂种克隆,包括两个保留人类17号染色体而无其他人类染色体的克隆,以检测人酸性α-葡萄糖苷酶和半乳糖激酶的表达。发现所有13个杂种克隆均表达这些人类酶。因此,我们得出结论,编码人酸性α-葡萄糖苷酶的基因位于人类17号染色体上。