Somatic cell hybridization of Roberts syndrome and normal human fibroblasts transfected with plasmids carrying dominant selection markers.
Somatic cell hybridization of Roberts syndrome and normal human fibroblasts transfected with plasmids carrying dominant selection markers.
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罗伯茨综合征和用携带显性选择标记的质粒转染的正常人成纤维细胞的体细胞杂交。
DOI：
10.1007/bf01535206
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Chang,PL
中科院分区：
文献类型：
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作者：
Gunby,JL;Tomkins,DJ;Chang,PL
罗伯茨综合征 (RS) 是一种罕见的人类隐性遗传疾病，某些患者的染色体会出现特征性的异染色质膨胀或分裂（RS 效应）。我们在具有异染色质异常的 RS 细胞株 (R22) 和具有正常染色体结构的次黄嘌呤磷酸核糖基转移酶缺陷的细胞株 (GM1662) 之间进行了体细胞杂交，以确定正常基因组的存在是否会纠正杂交细胞中的 RS 效应。为了向成纤维细胞菌株提供用于杂交的显性选择标记，用赋予抗生素G418抗性的质粒pSV3neo转染GM1662，并用赋予霉酚酸抗性的质粒pSV3gpt转染R22。进行两次体细胞杂交：(1)R22×GM1662 pSV3neo和(2)R22 pSVgpt×GM1662 pSV3neo。在每个实验中检查的 200 个杂交细胞中，分别有 95% 和 92% 的细胞不存在 RS 效应。这表明GM1662基因组能够纠正RS效应。一些杂交细胞中RS效应的存在归因于pSV3转染导致的不稳定核型，这可能导致这些杂交细胞中RS基因的正常等位基因的丢失。
Roberts syndrome (RS) is a rare human recessive disorder involving, in the chromosomes of some patients, a characteristic puffing or splitting apart of the constitutive heterochromatin (the RS effect). We carried out somatic cell hybridizations between an RS cell strain (R22) with the heterochromatin abnormality and a hypoxanthine phosphoribosyltransferase-deficient cell strain (GM1662) with normal chromosome structure to determine if the presence of the normal genome would correct the RS effect in the hybrid cells. In order to provide the fibroblast strains with dominant selection markers for the hybridizations, GM1662 was transfected with the plasmid pSV3neo which conferred resistance to the antibiotic G418, and R22 was transfected with the plasmid pSV3gpt which provided resistance to mycophenolic acid. Two somatic cell hybridizations were carried out: (1) R22×GM1662 pSV3neo and (2) R22 pSVgpt×GM1662 pSV3neo. The RS effect was found to be absent in 95% and 92%, respectively, of the 200 hybrid cells examined in each experiment. This indicated that the GM1662 genome was able to correct the RS effect. The presence of the RS effect in a few of the hybrid cells was attributed to the unstable karyotype resulting from pSV3 transfection which presumably caused the loss of the normal allele(s) of the RS gene in these hybrid cells.