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[在多药耐药的黑线仓鼠细胞中扩增的DNA序列的克隆与特性分析]

[Cloning and characteristics of DNA sequences amplified in Djungarian hamster cells with multidrug resistance].

作者信息

Chernova O B, Shifrin V I, Sokova O I, Kopnin B P, Gudkov A V

出版信息

Mol Gen Mikrobiol Virusol. 1987 Apr(4):14-9.

PMID:3600679
Abstract

A number of DNA clones containing the amplified DNA sequences were isolated from the genomic library of multidrug-resistant (MDR) Djungarian hamster cells using the DNAC0t 10-250 hybridization probe. Five independent nonoverlapping clones were obtained that covered more than 100 kb of the amplified genomic region. These clones were used as hybridization probes in blot-hybridization with DNA from 7 independently derived MDR Djungarian hamster cell lines selected for the resistance to colchicine or actinomycin D. Some clones contained the DNA sequences amplified in all of the cell lines tested while the others contained the cell line specific amplified sequences. Hybridization in situ was used to localize the amplified DNA in metaphase chromosomes of a MDR cell line that contained about 140 copies of these sequences. The approximate size of an amplicon calculated on the basis of the obtained data is about 1-2 X 10(3) kb.

摘要

使用DNA C0t 10 - 250杂交探针,从多药耐药(MDR)的西伯利亚仓鼠细胞基因组文库中分离出多个包含扩增DNA序列的DNA克隆。获得了五个独立的非重叠克隆,它们覆盖了超过100 kb的扩增基因组区域。这些克隆用作杂交探针,与从7个独立衍生的、对秋水仙碱或放线菌素D具有抗性的MDR西伯利亚仓鼠细胞系的DNA进行印迹杂交。一些克隆包含在所有测试细胞系中都扩增的DNA序列,而其他克隆则包含细胞系特异性的扩增序列。原位杂交用于将扩增的DNA定位到一个MDR细胞系的中期染色体上,该细胞系含有约140个这些序列的拷贝。根据获得的数据计算出的扩增子的大致大小约为1 - 2×10(3) kb。

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