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噬菌体AR1特异性感染大肠杆菌O157:H7,对其远端尾丝基因座进行表征并确定其受体。

Characterization of the distal tail fiber locus and determination of the receptor for phage AR1, which specifically infects Escherichia coli O157:H7.

作者信息

Yu S L, Ko K L, Chen C S, Chang Y C, Syu W J

机构信息

Institute of Microbiology and Immunology, National Yang Ming University, Pai-Tao, Taipei, 112, Taiwan.

出版信息

J Bacteriol. 2000 Nov;182(21):5962-8. doi: 10.1128/JB.182.21.5962-5968.2000.

Abstract

Phage AR1 is similar to phage T4 in several essential genes but differs in host range. AR1 infects various isolates of Escherichia coli O157:H7 but does not infect K-12 strains that are commonly infected by T4. We report here the determinants that confer this infection specificity. In T-even phages, gp37 and gp38 are components of the tail fiber that are critical for phage-host interaction. The counterparts in AR1 may be similarly important and, therefore, were characterized. The AR1 gp37 has a sequence that differs totally from those of T2 and T4, except for a short stretch at the N terminus. The gp38 sequence, however, has some conservation between AR1 and T2 but not between AR1 and T4. The sequences that are most closely related to the AR1 gp37 and gp38 are those of phage Ac3 in the T2 family. To identify the AR1-specific receptor, E. coli O157:H7 was mutated by Tn10 insertion and selected for an AR1-resistant phenotype. A mutant so obtained has an insertion occurring at ompC that encodes an outer membrane porin. To confirm the role of OmpC in the AR1 infection, homologous replacement was used to create an ompC disruption mutant (RM). When RM was complemented with OmpC originated from an O157:H7 strain, but not from K-12, its AR1 susceptibility was fully restored. Our results suggest that the host specificity of AR1 is mediated at least in part through the OmpC molecule.

摘要

噬菌体AR1在几个关键基因上与噬菌体T4相似,但宿主范围不同。AR1能感染多种大肠杆菌O157:H7分离株,但不能感染通常被T4感染的K-12菌株。我们在此报告赋予这种感染特异性的决定因素。在T偶数噬菌体中,gp37和gp38是尾丝的组成部分,对噬菌体与宿主的相互作用至关重要。AR1中的对应物可能同样重要,因此对其进行了表征。AR1的gp37除了N端有一小段外,其序列与T2和T4的序列完全不同。然而,gp38序列在AR1和T2之间有一些保守性,但在AR1和T4之间没有。与AR1的gp37和gp38最密切相关的序列是T2家族中噬菌体Ac3的序列。为了鉴定AR1特异性受体,通过Tn10插入对大肠杆菌O157:H7进行突变,并选择具有AR1抗性表型的菌株。如此获得的一个突变体在编码外膜孔蛋白的ompC处发生了插入。为了证实OmpC在AR1感染中的作用,使用同源替换创建了一个ompC破坏突变体(RM)。当RM用源自O157:H7菌株而非K-12菌株的OmpC进行互补时,其对AR1的敏感性完全恢复。我们的结果表明,AR1的宿主特异性至少部分是通过OmpC分子介导的。

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