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鼠RNA肿瘤病毒基因组的长片段代表性DNA拷贝的合成。

Synthesis of long, representative DNA copies of the murine RNA tumor virus genome.

作者信息

Rothenberg E, Baltimore D

出版信息

J Virol. 1975 Jan;17(1):168-74. doi: 10.1128/JVI.17.1.168-174.1976.

Abstract

Virions of Moloney murine leukemia virus can synthesize two classes of DNA molecules complementary to their 70S RNA. One class consists of molecules about 200 nucleotides long, which are of limited sequence complexity; these molecules are formed preferentially if the dNTP concentration during the reaction is low. The second class consists of very heterogeneous DNA molecules with weight-average size of about 1,000 nucleotides containing at least 70% of the viral RNA sequences in approximately equal concentration. The longest of these molecules can be 5,000 nucleotides long. This second class of DNA is formed in large amounts only in reactions containing dNTP concentrations of 0.2 mM or higher. In such reactions after 24 h of incubation, at least 35% of the input RNA is represented in DNA copies. The ability to make long, representative DNA transcripts of tumor virus RNA provides a source of excellent probes for molecular hybridization.

摘要

莫洛尼鼠白血病病毒的病毒粒子能够合成两类与它们的70S RNA互补的DNA分子。一类由长度约为200个核苷酸的分子组成,其序列复杂性有限;如果反应过程中的脱氧核苷酸三磷酸(dNTP)浓度较低,这些分子会优先形成。第二类由非常异质的DNA分子组成,重均大小约为1000个核苷酸,包含至少70%的病毒RNA序列,且浓度大致相等。这些分子中最长的可达5000个核苷酸长。只有在dNTP浓度为0.2 mM或更高的反应中,才会大量形成第二类DNA。在这样的反应中,孵育24小时后,至少35%的输入RNA会出现在DNA拷贝中。能够产生肿瘤病毒RNA的长的、具有代表性的DNA转录本,为分子杂交提供了优质探针来源。

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