大肠杆菌W3110的第7个IS1拷贝属于is1a (IS1E)型,这是唯一一个从染色体转位到质粒的IS1型。

J H Chen, H T Yeh
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摘要

在大肠杆菌K-12染色体中,已鉴定并鉴定了6个IS1拷贝(IS1A - IS1F)。根据其核苷酸(nt)序列,6个IS1拷贝可分为4种类型,IS1A(IS1E)、IS1B(IS1C)、IS1D和IS1F型。Zuber和Schumann(1993)在大肠杆菌W3110基因图谱上发现了第7个IS1拷贝,时间为49.6分钟。不幸的是,只有末端21-bp的序列和邻近的120-bp的大肠杆菌序列被报道。因此,我们设计了两个寡核苷酸引物,通过PCR特异性扩增第7个IS1拷贝。其中一个引物与IS1A(IS1E)、IS1B(IS1C)和IS1D的IRL序列的前16个碱基同源。另一个引物与第7个IS1拷贝的IRR相邻的大肠杆菌序列的18个碱基互补。获得一个800 bp的PCR片段并测定其nt序列,发现与IS1A(IS1E)的核苷酸序列相同。然后使用质粒系统分离由染色体插入序列插入引起的插入突变。在分离到的142个质粒插入突变体中,38个为IS5染色体插入,10个为IS30染色体插入,94个为IS1染色体插入。详细的酶切图谱显示,所有94个IS1插入的质粒都是IS1A(IS1E)型的IS1插入。
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The seventh copy of IS1 in Escherichia coli W3110 belongs to the IS1 A (IS1E) type which is the only IS1 type that transposes from chromosome to plasmids.

In the Escherichia coli K-12 chromosome, six copies of IS1 (IS1A - IS1F) have been identified and characterized. According to their nucleotide (nt) sequences, the six IS1 copies can be classified into four types, IS1A(IS1E), IS1B(IS1C), IS1D and IS1F type. Zuber and Schumann (1993) identified the seventh IS1 copy at 49.6 minutes on the E. coli W3110 genetic map. Unfortunately, only the end 21-bp sequence as well as the neighboring 120-bp E. coli sequence were reported. We therefore designed two oligonucleotide primers to specifically amplify the seventh IS1 copy by PCR. One primer is homologous to the first sixteen bases of the IRL sequence of IS1A(IS1E), IS1B(IS1C) and IS1D. The other primer is complementary to the eighteen bases of E. coli sequence adjacent to IRR of the seventh IS1 copy. An 800-bp PCR fragment was obtained and its nt sequence determined, revealing an identical nucleotide sequence to that of IS1A(IS1E). A plasmid system was then used to isolate insertion mutations caused by insertions of the chromosomal insertion sequences. Of the 142 plasmid insertion mutants isolated, thirty-eight were insertions of chromosomal IS5, ten were IS30, and ninety-four were IS1. Detailed restriction mapping indicates that all ninety-four plasmid IS1 insertions were insertions of IS1 of the IS1A(IS1E) type.

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