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整合多瘤病毒DNA的位点特异性切除

Site-specific excision of integrated polyoma DNA.

作者信息

Sylla B S, Huberdeau D, Bourgaux-Ramoisy D, Bourgaux P

出版信息

Cell. 1984 Jun;37(2):661-7. doi: 10.1016/0092-8674(84)90398-2.

Abstract

Cyp cells are permissive murine cells carrying a thermosensitive polyoma virus genome that remains integrated at 39 degrees C, but is effectively excised and replicated after transfer to 33 degrees C. In rare subclones of the Cyp line, temperature shift-down yields predominantly homogeneous populations of chimeric molecules that appear to reflect the circularization of defined segments of DNA spanning one of the junctions between the integrated viral genome and the adjacent cellular DNA. Such accurate and frequent excision requires a specific recombination mechanism. We examined both the cellular and the viral sequences that cross-over to generate one of these chimeric molecules, Rm I. The homology at the cross-over site is one of 1 or 2 base pairs at most; patches of homology, amounting in total to 19 or 20 base pairs, are found in perfect register on both sides of this site; and the two stretches of DNA that are joined to form RM I contain similar 12-14 base pair sequences (5'- CTCCTTTACAGAGG -3' and 5'- CTCCTTTCAAGG -3') in opposite orientations.

摘要

Cyp细胞是携带温度敏感型多瘤病毒基因组的允许性鼠细胞,该基因组在39℃时保持整合状态,但转移到33℃后会有效切除并复制。在Cyp细胞系的罕见亚克隆中,温度下调主要产生均匀的嵌合分子群体,这些分子似乎反映了跨越整合病毒基因组与相邻细胞DNA之间一个连接点的特定DNA片段的环化。这种精确且频繁的切除需要一种特定的重组机制。我们研究了产生这些嵌合分子之一Rm I的细胞和病毒序列。交叉位点的同源性最多为1或2个碱基对;在该位点两侧完美对齐的位置发现了总共19或20个碱基对的同源片段;连接形成RM I的两段DNA包含相反方向的相似的12 - 14个碱基对序列(5'- CTCCTTTACAGAGG -3'和5'- CTCCTTTCAAGG -3')。

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