Tischfield J A, Ruddle F H
Proc Natl Acad Sci U S A. 1974 Jan;71(1):45-9. doi: 10.1073/pnas.71.1.45.
A series of mouse-human hybrids was prepared from mouse cells deficient in adenine phosphoribosyltransferase (EC 2.4.2.7) and normal human cells. The hybrids were made in medium containing adenine and alanosine, an antimetabolite known to inhibit de novo adenylic acid biosynthesis. The mouse cells, unable to utilize exogenous adenine, were killed in this medium, but the hybrids proliferated as a consequence of their retaining the human aprt gene. The hybrids were then exposed to the adenine analogs 2,6-diaminopurine and 2-fluoroadenine to select for cells that had lost this gene. Before exposure to the adenine analogs, the expression of human adenine phosphoribosyltransferase by the hybrids was strongly associated only with the presence of human chromosome 16, and afterwards this was the only human chromosome consistently lost. This observation suggests that the human aprt gene can be assigned to chromosome 16.
从缺乏腺嘌呤磷酸核糖转移酶(EC 2.4.2.7)的小鼠细胞和正常人类细胞制备了一系列小鼠 - 人类杂种细胞。杂种细胞是在含有腺嘌呤和丙氨酸(一种已知可抑制从头合成腺苷酸生物合成的抗代谢物)的培养基中制备的。无法利用外源性腺嘌呤的小鼠细胞在此培养基中死亡,但杂种细胞由于保留了人类aprt基因而得以增殖。然后将杂种细胞暴露于腺嘌呤类似物2,6 - 二氨基嘌呤和2 - 氟腺嘌呤,以选择丢失该基因的细胞。在暴露于腺嘌呤类似物之前,杂种细胞中人类腺嘌呤磷酸核糖转移酶的表达仅与人类16号染色体的存在密切相关,而之后这是唯一持续丢失的人类染色体。这一观察结果表明,人类aprt基因可定位于16号染色体。