Alter B P, Goff S C, Hillman D G, Deisseroth A B, Forget B G
J Cell Sci. 1977 Aug;26:347-57. doi: 10.1242/jcs.26.1.347.
In an effort to activate the globin genes of non-erythroid cells, tetraploid murine erythroleukaemia cells (Friend cells) were fused with diploid human amniotic fibroblasts. When the Friend cells were pretreated with dimethylsulphoxide, an average of 27% heterokaryons was observed. These cells stained with benzidine, an indication that they contained haemoglobin. The cells incorporated radioactive amino acids into proteins. Electrophoresis of [3H]leucine-labelled lysates on SDS urea polyacrylamide gels indicated that up to 7% of the newly synthesized protein co-electrophoresed with globin. CM cellulose chromatography demonstrated the presence of mouse but not human globin chains. Hybridization analyses of cytoplasmic RNA also revealed only mouse globin mRNA in the heterokaryons. Although heterokaryons form readily between mouse erythroleukaemia cells and human fibroblasts, and globin synthesis does occur, only the erythroid partner in the fusion system employed here directs globin production.
为了激活非红细胞系细胞的珠蛋白基因，将四倍体小鼠红白血病细胞（弗氏细胞）与二倍体人羊膜成纤维细胞进行融合。当用二甲亚砜预处理弗氏细胞时，观察到平均27%的异核体。这些细胞用联苯胺染色，表明它们含有血红蛋白。这些细胞将放射性氨基酸掺入蛋白质中。在十二烷基硫酸钠 - 尿素聚丙烯酰胺凝胶上对[³H]亮氨酸标记的裂解物进行电泳表明，高达7%的新合成蛋白质与珠蛋白共电泳。羧甲基纤维素层析证明存在小鼠珠蛋白链，但不存在人珠蛋白链。对细胞质RNA的杂交分析也显示异核体中仅存在小鼠珠蛋白mRNA。尽管小鼠红白血病细胞与人成纤维细胞之间很容易形成异核体，并且确实发生了珠蛋白合成，但在此处使用的融合系统中，只有红细胞系伙伴指导珠蛋白的产生。
