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用于定量大肠杆菌中Tn5倒位事件的简易测定法及其在测定质粒拷贝数中的应用。

Simple assay for quantitation of Tn5 inversion events in Escherichia coli and use of the assay in determination of plasmid copy number.

作者信息

Weber P C, Levine M, Glorioso J C

机构信息

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109.

出版信息

J Bacteriol. 1988 Oct;170(10):4972-5. doi: 10.1128/jb.170.10.4972-4975.1988.

Abstract

In this report, we describe a simple method for measuring the frequency of sequence inversion in the transposable element Tn5 as a result of recombination across its duplicated IS50 elements. The structure of Tn5 was manipulated so that the neomycin phosphotransferase gene of the transposon would be expressed only if a sequence inversion event occurred. This highly sensitive assay also served as the basis for a novel means of estimating plasmid copy number.

摘要

在本报告中,我们描述了一种简单的方法,用于测量转座元件Tn5中由于其重复的IS50元件间的重组而导致的序列倒位频率。对Tn5的结构进行了操作,使得只有在发生序列倒位事件时转座子的新霉素磷酸转移酶基因才会表达。这种高度灵敏的检测方法还为估算质粒拷贝数的新方法奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/08d1/211551/6ba70dca23ea/jbacter00188-0555-a.jpg

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