Cloning of DNA from double minutes of Y1 mouse adrenocortical tumor cells: Evidence for gene amplification
Cloning of DNA from double minutes of Y1 mouse adrenocortical tumor cells: Evidence for gene amplification
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从双分钟 Y1 小鼠肾上腺皮质肿瘤细胞中克隆 DNA：基因扩增的证据
DOI：
10.1016/0092-8674(81)90507-9
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发表时间：
1981
期刊：
影响因子：
64.5
通讯作者：
V. E. Powers
中科院分区：
文献类型：
--
作者：
D. George;V. E. Powers
我们从小鼠肾上腺皮质肿瘤细胞系（Yl-DM）中分离出一个高度富集双微体（DM）的中期染色体片段。我们已经克隆了DNA从这个DM-丰富的馏分中的X载体Charon 4A，并具有其特征在于两个随机选择的重组噬菌体克隆从这个DM DNA文库。当32 P标记的DNA从每个重组体杂交到Southern印迹的限制性内切酶消化的DNA从不同的小鼠细胞系，很大的差异被认为是在所得的放射自显影图像的强度，这取决于基因组DNA的来源。用来自Y1-DM细胞的DNA和来自相关Y1亚系的DNA获得非常强的信号，所述相关Y1亚系缺乏dm但含有带有大的均匀染色区域（HSR）的标记染色体。与来自亲本近交系小鼠和来自两个不相关的小鼠细胞系的DNA杂交产生的信号比用来自Y1细胞的DNA获得的信号显著更弱，但是来自这些来源的DNA片段具有相似的大小。基于过滤杂交分析的结果，我们估计与克隆片段同源的序列在Y1-DM细胞的基因组中比在亲本小鼠细胞中丰度高约1DD-至2 OD-倍。这些数据与这些细胞中的dm和HSR含有扩增基因的假设一致。
We have isolated a metaphase chromosome fraction highly enriched in double minutes (dm) from a mouse adrenocortical tumor cell line (Yl-DM). We have cloned DNA from this dm-enriched fraction in the X vector Charon 4A, and have characterized two randomly chosen recombinant bacteriophage clones from this dm DNA library. When 32P-labeled DNA from each recombinant was hybridized to Southern blots of restriction endonuclease-digested DNA from different mouse cell lines, large differences were seen in the intensity of the resulting autoradiographic images, depending on the source of the genomic DNA. A very strong signal was obtained with DNA from the Yl-DM cells and with DNA from a related Yl subline that lacks dm but contains a marker chromosome bearing a large homogeneously staining region (HSR). Hybridization to DNA from parental inbred mice and from two unrelated mouse cell lines produced a significantly weaker signal than that obtained with DNA from the Yl cells, but the DNA fragments from these sources were of similar size. Based on results from filter hybridization analysis, we estimate that sequences homologous to the cloned fragments are approximately lDD-to 2OD-fold more abundant in the genome of the Yl-DM cells than in the parental mouse cells. The data are consistent with the hypothesis that dm and HSRs in these cells contain amplified genes.
对小鼠肾上腺皮质肿瘤细胞系中的染色体区域和双分钟进行均匀染色。
DOI：
10.1159/000131535
发表时间：
1980
期刊：
Cytogenetics and cell genetics
影响因子：
--
作者：
George,DL;Francke,U
通讯作者：
Francke,U