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葡萄球菌噬菌体L54a通过位点特异性重组实现整合:附着位点的结构分析

Integration of staphylococcal phage L54a occurs by site-specific recombination: structural analysis of the attachment sites.

作者信息

Lee C Y, Iandolo J J

出版信息

Proc Natl Acad Sci U S A. 1986 Aug;83(15):5474-8. doi: 10.1073/pnas.83.15.5474.

Abstract

Lysogenization by staphylococcal phage L54a induces the loss of lipase (glycerol ester hydrolase) activity in its host Staphylococcus aureus. The attachment site of the bacterial chromosome (attB) for the phage is at the 3' end of the lipase gene, geh. The DNA fragment containing the attB (base pairs 2620-2637 inclusive) site has been sequenced. We have also cloned and determined the nucleotide sequence of the DNA fragments containing the other three attachment sites--i.e., the attP locus on the circularly permuted phage genome and the attL and attR loci at the left and right ends of the prophage in the lysogenized strain. These results reveal that an 18-base-pair core sequence is common to all four att sites. These data indicate that the crossover point must exist within the core sequence and, further, that integration is site- and orientation-specific. We also localized the viral recombinase gene to a 2.1-kilobase DNA segment extending rightward to the attP site. This region was found to be essential for integration of plasmids containing the attP site.

摘要

葡萄球菌噬菌体L54a的溶原化作用会导致其宿主金黄色葡萄球菌的脂肪酶(甘油酯水解酶)活性丧失。该噬菌体在细菌染色体上的附着位点(attB)位于脂肪酶基因geh的3'端。已对包含attB位点(碱基对2620 - 2637,含两端)的DNA片段进行了测序。我们还克隆并测定了包含其他三个附着位点的DNA片段的核苷酸序列,即环状排列的噬菌体基因组上的attP位点以及溶原化菌株中前噬菌体左右两端的attL和attR位点。这些结果表明,一个18碱基对的核心序列在所有四个att位点中是共有的。这些数据表明交叉点必定存在于核心序列内,而且进一步表明整合是位点特异性和方向特异性的。我们还将病毒重组酶基因定位到一个2.1千碱基的DNA片段,该片段向右延伸至attP位点。发现该区域对于含有attP位点的质粒的整合至关重要。

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