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由合成的IS50(Tn5)衍生物进行的分子内转座。

Intramolecular transposition by a synthetic IS50 (Tn5) derivative.

作者信息

Tomcsanyi T, Berg C M, Phadnis S H, Berg D E

机构信息

Department of Molecular Microbiology, Washington University Medical School, St. Louis, Missouri 63110-1093.

出版信息

J Bacteriol. 1990 Nov;172(11):6348-54. doi: 10.1128/jb.172.11.6348-6354.1990.

Abstract

We report the formation of deletions and inversions by intramolecular transposition of Tn5-derived mobile elements. The synthetic transposons used contained the IS50 O and I end segments and the transposase gene, a contraselectable gene encoding sucrose sensitivity (sacB), antibiotic resistance genes, and a plasmid replication origin. Both deletions and inversions were associated with loss of a 300-bp segment that is designated the vector because it is outside of the transposon. Deletions were severalfold more frequent than inversions, perhaps reflecting constraints on DNA twisting or abortive transposition. Restriction and DNA sequence analyses showed that both types of rearrangements extended from one transposon end to many different sites in target DNA. In the case of inversions, transposition generated 9-bp direct repeats of target sequences.

摘要

我们报道了通过Tn5衍生的移动元件的分子内转座形成缺失和倒位的情况。所使用的合成转座子包含IS50 O和I末端片段以及转座酶基因、编码蔗糖敏感性的反选择基因(sacB)、抗生素抗性基因和质粒复制起点。缺失和倒位都与一个300碱基对片段的丢失有关,该片段被称为载体,因为它位于转座子之外。缺失的发生频率比倒位高几倍,这可能反映了对DNA扭曲或流产转座的限制。限制性酶切和DNA序列分析表明,这两种类型的重排都从一个转座子末端延伸到靶DNA中的许多不同位点。在倒位的情况下,转座产生了靶序列的9碱基对直接重复序列。

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