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Tn5对称反向重复序列的精确及近乎精确切除;大肠杆菌中不依赖recA的缺失事件的共同特征

Precise and nearly-precise excision of the symmetrical inverted repeats of Tn5; common features of recA-independent deletion events in Escherichia coli.

作者信息

Collins J, Volckaert G, Nevers P

出版信息

Gene. 1982 Jul-Aug;19(1):139-46. doi: 10.1016/0378-1119(82)90198-6.

Abstract

The transposon Tn5 contains a unique central region bordered by 1.5-kb inverted repeats. The in vitro deletion of the centre of Tn5, with a restriction endonuclease (XhoI) which cuts within the inverted repeats leads to the production of a palindrome on subsequent ligation. This palindromic region is unstable on subsequent transformation into Escherichia coli (Collins, 1981). Precise excision of the Tn5 region plus one copy of the bracketing 9-bp direct repeat occurred in about one-third of the transformants. The rest of the transformants contain only remnants of the inverted repeat. Sequence analysis indicated that deletion had occurred between short direct repeats. The precise excision of these "nearly precise" excision products continued with high frequency and was found to be affected by mutations that interfere with the normal precise excision of transposons. In a recB, sbcB host precise excision was markedly reduced. A common mechanism is proposed for all recA-independent deletions occurring in E. coli.

摘要

转座子Tn5包含一个独特的中央区域,其两侧为1.5kb的反向重复序列。用一种在反向重复序列内切割的限制性内切酶(XhoI)对Tn5的中心进行体外缺失,随后连接会产生一个回文结构。这个回文区域在随后转化到大肠杆菌中时是不稳定的(柯林斯,1981)。在大约三分之一的转化子中发生了Tn5区域加上一份侧翼9bp直接重复序列的精确切除。其余的转化子只含有反向重复序列的残余部分。序列分析表明,缺失发生在短的直接重复序列之间。这些“近乎精确”切除产物的精确切除以高频率持续进行,并且发现受到干扰转座子正常精确切除的突变的影响。在recB、sbcB宿主中,精确切除明显减少。针对大肠杆菌中所有不依赖recA的缺失提出了一种共同机制。

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