McBride O W, Ozer H L
Proc Natl Acad Sci U S A. 1973 Apr;70(4):1258-62. doi: 10.1073/pnas.70.4.1258.
Transfer of genetic information from isolated mammalian chromosomes to recipient cells has been demonstrated. Metaphase chromosomes isolated from Chinese hamster fibroblasts were incubated with mouse A(9) cells containing a mutation at the hypoxanthine-guanine phosphoribosyl transferase (hprt) locus. Cells were plated in a selective medium, resulting in death of all unaltered parental A(9) cells. However, colonies of cells containing hypoxanthine phosphoribosyl transferase (EC 2.4.2.8) appeared with a variable frequency of about 10(-6) to 10(-7). The enzyme from these cells was indistinguishable from that from Chinese hamster cells, as shown by DEAE-cellulose chromatography and gel electrophoresis, and differed clearly from the mouse enzyme. The colonies, thus, did not result from reversion of A(9) parental cells to wild type, but appeared to represent progeny of individual cells that had ingested chromosomes, replicated, and expressed the hprt gene. These colonies differed from each other in stability of expression of the transferred gene.
遗传信息从分离的哺乳动物染色体转移到受体细胞已得到证实。从中国仓鼠成纤维细胞分离的中期染色体与在次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(hprt)基因座处有突变的小鼠A(9)细胞一起孵育。将细胞接种在选择性培养基中,导致所有未改变的亲代A(9)细胞死亡。然而,含有次黄嘌呤磷酸核糖转移酶(EC 2.4.2.8)的细胞集落以约10^(-6)至10^(-7)的可变频率出现。通过DEAE - 纤维素色谱法和凝胶电泳显示,这些细胞中的酶与中国仓鼠细胞中的酶无法区分,并且与小鼠酶明显不同。因此,这些集落不是由A(9)亲代细胞回复为野生型产生的,而是似乎代表摄取了染色体、进行了复制并表达了hprt基因的单个细胞的后代。这些集落在转移基因表达的稳定性方面彼此不同。