Francke U, Busby N, Shaw D, Hansen S, Brown M G
Somatic Cell Genet. 1976 Jan;2(1):27-40. doi: 10.1007/BF01539240.
The structural gene for purine-nucleoside phosphorylase (NP) has been assigned to a subregion of chromosome 14 by somatic cell hybridization of male and female cells containing the balanced translocation t(X;14) (p22;q21). Peripheral lymphocytes were fused to a pseudodiploid HPRT-deficient established Chinese hamster cell line. 23 primary hybrid clones (10 derived from male and 13 from female cells) were isolated and maintained in HAT selective medium. Parallel subcultures from generations 16, 24, and 40 after clonal isolation were fully karyotyped and analyzed electrophorectically for expression of the human types of NP, HPRT, G6PD, and PGK. The human NP phenotype segregated discordantly with each human chromosome except chromosome 14 and the der(14),t(X;14) translocation chromosome. In all, 8 hybrids which had retained the der(X), t(X;14) translocation chromosome under HAT selective pressure and expressed human HPRT had lost the human NP phenotype. These results indicate localization of the NP gene in region 14pter leads to 14q21.
通过对含有平衡易位t(X;14)(p22;q21)的雄性和雌性细胞进行体细胞杂交，嘌呤核苷磷酸化酶（NP）的结构基因已被定位到14号染色体的一个亚区域。外周淋巴细胞与一个伪二倍体、缺乏次黄嘌呤磷酸核糖转移酶（HPRT）的中国仓鼠细胞系进行融合。分离出23个初级杂交克隆（10个来自雄性细胞，13个来自雌性细胞），并在次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷（HAT）选择培养基中进行培养。对克隆分离后第16、24和40代的平行传代培养物进行了完整的核型分析，并通过电泳分析了人类NP、HPRT、葡萄糖-6-磷酸脱氢酶（G6PD）和磷酸甘油酸激酶（PGK）的表达情况。人类NP表型与除14号染色体和der(14)、t(X;14)易位染色体之外的每一条人类染色体均呈不一致分离。在HAT选择压力下，共有8个保留了der(X)、t(X;14)易位染色体并表达人类HPRT的杂交克隆失去了人类NP表型。这些结果表明NP基因定位于14号染色体的14pter至14q21区域。
