Replicon fusions promoted by the inverted repeats of Tn5. The right repeat is an insertion sequence.

Replicon fusions promoted by the inverted repeats of Tn5. The right repeat is an insertion sequence.
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Tn5 的反向重复序列促进复制子融合。

DOI:
10.1016/0022-2836(81)90322-3
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发表时间:
1981
影响因子:
5.6
通讯作者:
Syvanen,M
Syvanen,M
中科院分区:
生物学2区
文献类型:
--
作者:
Isberg,RR;Syvanen,M

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在大肠杆菌中,有几种转座因子能够促进复制子融合。我们在这里展示了转座子Tn 5也具有这种性质。Tn 5能够以Tn 5转座到λ上的频率的2%促进质粒pBR 322(其所驻留的)与λ噬菌体之间的融合。在完全缺乏同源重组和整合重组的菌株中形成的质粒/噬菌体复制子融合体总是由λ和pBR 322连接处的插入元件的直接复制组成。为了确定Tn 5中引起复制子融合所必需的区域,在体外和体内构建了一系列Tn 5缺失。删除衍生物,删除所有,但1500个碱基对的右倒重复(茎)是有能力促进融合。左反向重复不能提供这种活性,但可以被反式互补以形成复制子融合体。我们的结论是,两个重复序列都具有复制子融合所必需的位点,但只有正确的茎具有所需的蛋白质。这确定了正确的重复序列是插入序列。
Several transposable elements are able to promote replicon fusions inEscherichia coli. We show here that the transposon, Tn5, also has this property. Tn5 is able to promote fusions between the plasmid pBR322 (on which it resides) and bacteriophage λ at 2% of the frequency of Tn5 transposition onto λ. Plasmid/phage replicon fusions formed in strains that are totally deficient in homologous and integrative recombination always consist of direct duplications of the insertion element at the junctions between λ and pBR322.In order to define those regions within the Tn5 necessary for causing replicon fusions, a series of deletions of Tn5 generatedin vitroandin vivowere constructed. Deletion derivatives that remove all but the 1500-base-pair right inverted repetition (stem) are competent to promote fusions. The left inverted repetition is unable to supply this activity, but can be complemented intransto form replicon fusions. We conclude that both repeats have the sites necessary for replicon fusions, but only the right stem has the required protein(s). This establishes that the right repeat is an insertion sequence.