Eun C K, Paik S G, Goldwasser P, Shin S, Klinger H P
Cytogenet Cell Genet. 1981;29(2):116-21. doi: 10.1159/000131558.
Immunochemical methods were used to identify the genetic origin of hypoxanthine phosphoribosyltransferase (HPRT) expressed in heteroploid, HPRT-deficient mouse (A9) cells and Chinese hamster ovary (K627) cells, after these cells were fused with chick embryo erythrocytes and selected for resistance to hypoxanthine-aminopterin-thymidine (HAT) medium. All of the HAT-selected clones produced HPRT activity which was immunoprecipitable by an antiserum specific for chick HPRT, but not by an antiserum specific for mouse and hamster HPRT. Furthermore, the HPRT activity in these clones was electrophoretically indistinguishable from chick liver HPRT and clearly different from mouse liver HPRT. These data provide evidence that the HPRT activity expressed in cell hybrids produced by the fusion of HPRT-negative mammalian cells and chick erythrocytes containing genetically inactive nuclei is indeed coded by the chick HPRT gene and that an avian gene can be stably incorporated and correctly expressed in a mammalian cells.
采用免疫化学方法鉴定次黄嘌呤磷酸核糖转移酶(HPRT)的基因来源,该酶在异倍体、HPRT缺陷型小鼠(A9)细胞和中国仓鼠卵巢(K627)细胞中表达,这些细胞与鸡胚红细胞融合后,筛选出对次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷(HAT)培养基具有抗性的细胞。所有经HAT筛选的克隆均产生HPRT活性,该活性可被针对鸡HPRT的抗血清免疫沉淀,但不能被针对小鼠和仓鼠HPRT的抗血清免疫沉淀。此外,这些克隆中的HPRT活性在电泳上与鸡肝HPRT无法区分,与小鼠肝HPRT明显不同。这些数据表明,由HPRT阴性哺乳动物细胞与含有遗传失活细胞核的鸡红细胞融合产生的细胞杂种中表达的HPRT活性确实由鸡HPRT基因编码,并且禽类基因可以稳定整合并在哺乳动物细胞中正确表达。