甲氨蝶呤抗性小鼠细胞大标记染色体中的卫星DNA
Satellite DNA in large marker chromosomes of methotrexate-resistant mouse cells.
作者信息
Bostock C J, Clark E M
出版信息
Cell. 1980 Mar;19(3):709-15. doi: 10.1016/s0092-8674(80)80047-x.
PMID:7363327
Abstract
The additional segments of five large marker chromosomes of methotrexate-resistant mouse melanoma PG19T3 cells are shown to consist of C-banding material. In situ hybridization indicates that these additional segments also contain a high proportion of sequences that will cross hybridize with mouse satellite DNA. Analytical density gradient centrifugation suggests that up to 60% of the DNA in the additional segments may be in the form of satellite DNA. Incorporation of bromodeoxyuridine for one complete S phase and staining with the Hoechst 33258 fluorescence plus Giemsa technique reveals complex asymmetries within the additional segments. These asymmetries are interpreted as showing a large repeating unit, which is likely to be involved in the amplification of the dihydrofolate reductase gene.
摘要
甲氨蝶呤耐药小鼠黑色素瘤PG19T3细胞的五条大型标记染色体的额外片段显示由C带物质组成。原位杂交表明，这些额外片段还含有高比例的能与小鼠卫星DNA交叉杂交的序列。分析性密度梯度离心表明，额外片段中高达60%的DNA可能是卫星DNA的形式。用溴脱氧尿苷掺入一个完整的S期，并采用Hoechst 33258荧光加吉姆萨技术染色，揭示了额外片段内复杂的不对称性。这些不对称性被解释为显示出一个大的重复单元，它可能参与二氢叶酸还原酶基因的扩增。
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