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天蓝色链霉菌中介导SLP1元件位点特异性重组的基因的定位和核苷酸序列。

Localization and nucleotide sequences of genes mediating site-specific recombination of the SLP1 element in Streptomyces lividans.

作者信息

Brasch M A, Pettis G S, Lee S C, Cohen S N

机构信息

Department of Genetics, Stanford University School of Medicine, California 94305.

出版信息

J Bacteriol. 1993 May;175(10):3067-74. doi: 10.1128/jb.175.10.3067-3074.1993.

Abstract

SLP1 is a 17.2-kbp genetic element indigenous to the Streptomyces coelicolor chromosome. During conjugation, SLP1 can undergo excision and subsequent site-specific integration into the chromosomes of recipient cells. We report here the localization, nucleotide sequences, and initial characterization of the genes mediating these recombination events. A region of SLP1 adjacent to the previously identified site of integration, attP, was found to be sufficient to promote site-specific integration of an unrelated Streptomyces plasmid. Nucleotide sequence analysis of a 2.2-kb segment of this region reveals two open reading frames that are adjacent to and transcribed toward the attP site. One of these, the 1,365-bp int gene of SLP1, encodes a predicted 50.6-kDa basic protein having substantial amino acid sequence similarity to a family of site-specific recombinases that includes the Escherichia coli bacteriophage lambda integrase. A linker insertion in the 5' end of the cloned int gene prevents integration, indicating that Int is essential for promoting integration. An open reading frame (orf61) lying immediately 5' to int encodes a predicted 7.1-kDa basic peptide showing limited sequence similarity to the excisionase (xis) genes of other site-specific recombination systems.

摘要

SLP1是天蓝色链霉菌染色体特有的一个17.2千碱基对的遗传元件。在接合过程中,SLP1可进行切除并随后位点特异性整合到受体细胞的染色体中。我们在此报告介导这些重组事件的基因的定位、核苷酸序列及初步特性。发现SLP1中与先前鉴定的整合位点attP相邻的一个区域足以促进无关链霉菌质粒的位点特异性整合。对该区域一个2.2千碱基片段的核苷酸序列分析揭示了两个与attP位点相邻并朝该位点转录的开放阅读框。其中之一,即SLP1的1365碱基对int基因,编码一种预测分子量为50.6千道尔顿的碱性蛋白,该蛋白与包括大肠杆菌噬菌体λ整合酶在内的一个位点特异性重组酶家族具有显著的氨基酸序列相似性。克隆的int基因5′端的一个接头插入会阻止整合,这表明Int对于促进整合至关重要。紧挨着int 5′端的一个开放阅读框(orf61)编码一种预测分子量为7.1千道尔顿的碱性肽,该肽与其他位点特异性重组系统的切除酶(xis)基因显示出有限的序列相似性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/486c/204627/892f75f1565b/jbacter00052-0287-a.jpg

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