CHROMOSOME ASSIGNMENT OF A MURINE GLUCOCORTICOID RECEPTOR GENE (GRL-1) USING INTRASPECIES SOMATIC-CELL HYBRIDS
CHROMOSOME ASSIGNMENT OF A MURINE GLUCOCORTICOID RECEPTOR GENE (GRL-1) USING INTRASPECIES SOMATIC-CELL HYBRIDS
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DOI：
10.1016/0092-8674(80)90541-3
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发表时间：
1980-01-01
期刊：
影响因子：
64.5
通讯作者：
GEHRING, U
中科院分区：
文献类型：
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作者：
FRANCKE, U;GEHRING, U
2个小鼠淋巴瘤细胞系，糖皮质激素敏感的S49.1和耐药的EL 4系之间的杂交，是敏感的溶细胞类固醇的作用，只要他们保留S49.1特异性糖皮质激素受体和完整的染色体互补贡献的双亲线。通过使用半合成糖皮质激素地塞米松，分离出失去S49.1特异性受体但保留EL 4型受体的抗性分离体（dexR）。平均而言，这些分离子也丢失了1条染色体。用胰蛋白酶-姬姆萨显带法对亲本细胞系及其杂种进行了染色体核型分析。S49.1细胞含有40条明显正常的染色体，具有X单体和1三体。EL 4细胞有39条染色体，其中一半结构异常。除了在S49.1 × 10中低水平的随机核型变异性之外，EL 4杂交种中，在敏感（dexS）和dexR亚系之间观察到1个一致的差异：敏感杂交种含有3条18号染色体，而抗性分离体只有2条。由S49.1亲本贡献的18号染色体与由EL 4细胞系贡献的染色体通过涉及着丝粒异染色质和核仁组织者区的结构变异而可区分。来自S49.1的一条特异的18号染色体在来自不同dexS杂种无性系的3个dexR分离系中始终缺失。由于dexR分离子杂交体已经丢失了S49.1特异性糖皮质激素受体，该受体的基因显然位于始终丢失的18号染色体上。其他S49.1衍生的18可能携带突变或沉默受体基因，因为S49/1先前已显示具有半合子水平的受体。强调了种内体细胞杂种用于作图的有用性。这种杂种的总体核型稳定性允许鉴定被选择压力消除的特定染色体。
Hybrids between 2 mouse lymphoma cell lines, the glucocorticoid sensitive S49.1 and the resistant EL4 line, are sensitive to the cytolytic steroid effect as long as they retain the S49.1 specific glucocorticoid receptor and the complete chromosome complements contributed by both parental lines. With the use of the semisynthetic glucocorticoid dexamethasone, resistant segregants (dexR) were isolated which have lost the S49.1 specific receptor but retain the EL4 type of receptor. On the average, these segregants have also lost 1 chromosome. A detailed karyotype analysis of both parental cell lines and their hybrids hybrids was carried out using trypsin-Giemsa banding for chromosome identification. S49.1 cells contain 40 apparently normal chromosomes with monosomy X and trisomy 1. EL4 cells have 39 chromosomes, half of which are structurally abnormal. In addition to low levels of random karyotypic variability in S49.1 .times. EL4 hybrids, 1 consistent difference was observed between sensitive (dexS) and dexR sublines: sensitive hybrids contained 3 chromosomes 18 while resistant segregants only had 2. The chromosome 18 contributed by the S49.1 parent was distinguishable from the one contributed by the EL4 cell line by structural variations involving the centromeric heterochromatin and the nucleolar organizer regions. A specific chromosome 18 derived from S49.1 was consistently absent in 3 dexR segregant lines derived from different dexS hybrid clones. Since the dexR segregant hybrids have lost the S49.1 specific glucocorticoid receptor, the gene for the receptor is apparently located on that chromosome 18 which is consistently lost. The other S49.1 derived 18 presumably carries a mutant or silent receptor gene, as S49/1 has previously been shown to have hemizygous levels of the receptor. The usefulness of intraspecies somatic cell hybrids for mapping purposes is emphasized. The overall karyotypic stability of such hybrids allows the identification of specific chromosomes eliminated by selection pressure.