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一种检测异源DNA之间同源性并进行图谱绘制的快速方法。多瘤病毒基因组的评估。

A rapid method for detecting and mapping homology between heterologous DNAs. Evaluation of polyomavirus genomes.

作者信息

Howley P M, Israel M A, Law M F, Martin M A

出版信息

J Biol Chem. 1979 Jun 10;254(11):4876-83.

PMID:220264
Abstract

A new approach for evaluating homologous sequences among related DNAs is presented. Conventional filter hybridization techniques are employed at 35 degrees C in a range of formamide concentrations in order to perform annealings at effective temperatures as low as Tm -50 degrees C which permits the detection of regions of homology with as much as 33% base mismatch. Under such nonstringent conditions, high levels of specific annealing can be obtained at plateau levels. In combination with the Southern "blotting" technique (1975), this approach can be used to perform biochemical heteroduplex melting experiments. The homology among the genomes of the murine polyoma virus (Py), the simian virus 40 (SV40), and the human papovavirus BK was evaluated using this new methodology.

摘要

本文提出了一种评估相关DNA之间同源序列的新方法。在一系列甲酰胺浓度下于35℃采用传统的滤膜杂交技术,以便在低至Tm - 50℃的有效温度下进行退火,这允许检测具有高达33%碱基错配的同源区域。在这种非严格条件下,可在平台水平获得高水平的特异性退火。结合Southern“印迹”技术(1975年),该方法可用于进行生化异源双链体解链实验。使用这种新方法评估了小鼠多瘤病毒(Py)、猴病毒40(SV40)和人乳头瘤病毒BK的基因组之间的同源性。

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