Resolvase-like serine recombinase mediates integration/excision in the bacteriophage φRSM
Resolvase-like serine recombinase mediates integration/excision in the bacteriophage φRSM
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DOI：
10.1016/j.jbiosc.2010.10.001
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发表时间：
2011-02-01
影响因子：
2.8
通讯作者：
Yamada, Takashi
中科院分区：
文献类型：
--
作者：
Askora, Ahmed;Kawasaki, Takeru;Yamada, Takashi
感染青枯雷尔氏菌（Ralstonia solanacearum）的丝状噬菌体phi RSM 1编码与位点特异性丝氨酸重组酶的解离酶/转化酶亚家族成员具有显著同源性的开放阅读框（ORF 14）。类似的前噬菌体存在于各种R.青枯菌R. pickettii和类鼻疽伯克霍尔德氏菌。我们的特点是整合和切除重组反应介导的phi RSM 1整合酶使用体内试验。大肠结果表明，含有RSM 1 orf 14和attP序列的大肠杆菌质粒（pT-orf 14-attP）整合到R.青枯菌细胞pT-orf 14-attP和pS-attB（a R.含有attB的青枯菌质粒）也存在于R.青枯菌细胞attP/attB重组的产物，即，attL和attR与青枯菌菌株中原噬菌体phi RSM中发现的序列完全相同。pRecomb质粒上attL和attR序列之间的分子内重组（切除重组）也显示以orf 14依赖性方式发生。这是第一个证据表明，一个小的丝氨酸重组酶从分解酶/转化酶组可能功能的整合和切除重组丝状真菌。(C)2010年，生物技术学会，日本。All rights reserved.
phi RSM1, a filamentous phage infecting Ralstonia solanacearum, encodes an open reading frame (ORF14) that exhibits significant homology to members of the resolvase/invertase subfamily of site-specific serine recombinases. Similar prophages are found in the genomes of various strains of R. solanacearum, R. pickettii, and Burkholderia pseudomallei. We have characterized the integrative and excisive recombination reactions mediated by the phi RSM1 integrase using in vivo assays. An E. coli plasmid containing the RSM1 orf14 and attP sequences (pT-orf14-attP) was shown to integrate into the attB sequence of R. solanacearum cells. Intermolecular recombination between pT-orf14-attP and pS-attB (a R. solanacearum plasmid containing attB) also occurred in R. solanacearum cells. The products of attP/attB recombination, i.e., attL and attR, were exactly identical to the sequences found in the prophage phi RSM in it solanacearum strains. Intramolecular recombination between the attL and attR sequences (excisive recombination) on the pRecomb plasmid was also shown to occur in an orf14-dependent manner. This is the first evidence that a small serine recombinase from the resolvase/invertase group may function in integrative and excisive recombination for filamentous phages. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.