Characterization of a novel T4-type Stenotrophomonas maltophilia virulent phage Smp14
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S H O R T CO M MU N I C A T I O N
Characterization of a novel T4-type Stenotrophomonas maltophilia virulent phage Smp14 Chiy-Rong Chen · Ching-Hsuan Lin · Juey-Wen Lin · Chi-I Chang · Yi-Hsiung Tseng · Shu-Fen Weng
Received: 5 February 2007 / Revised: 19 March 2007 / Accepted: 24 March 2007 / Published online: 18 April 2007 © Springer-Verlag 2007
Abstract Stenotrophomonas maltophilia (Sm), with most of the isolates being resistant to multidrugs, is an opportunistic bacterium causing nosocomial infections. In this study, a novel virulent Sm phage, Smp14, was characterized. Electron microscopy showed that Smp14 resembled members of Myoviridae and adsorbed to poles of the host cells during infection. It lysed 37 of 87 clinical Sm isolates in spot test, displayed a latent period of ca. 20 min, and had a burst size of ca. 150. Its genome (estimated to be 160 kb by PFGE), containing m4C and two unknown modiWed bases other than m5C and m6A as identiWed by HPLC, resisted to digestion with many restriction endonucleases except MseI. These properties indicate that it is a novel Sm
phage distinct from the previously reported phiSMA5 which has a genome of 250 kb digestible with various restriction enzymes. Sequencing of a 16 kb region revealed 12 ORFs encoding structural proteins sharing 15–45% identities with the homologues from T4-type phages. SDSPAGE displayed 20 virion proteins, with the most abundant one being the 39 kDa major capsid protein (gp23), which had the N-terminal 52 amino acids removed. Phylogenetic analysis based on gp23 classiWed Smp14 into a novel single-membered T4-type subgroup. Keywords S. maltophilia · Virulent phage · T4-type · Unknown modiWed bases · Major capsid protein · Phylogenetic analysis
Communicated by Jorge Membrillo-Hernández. Chiy-Rong Chen and Ching-Hsuan Lin contributed equally. Electronic supplementary material The online version of this article (doi:10.1007/s00203-007-0238-5) contains supplementary material, which is available to authorized users. C.-R. Chen · C.-H. Lin · S.-F. Weng (&) Institute of Molecular Biology, National Chung Hsing University, Taichung, 402, Taiwan e-mail: [email protected] J.-W. Lin Institute of Biochemistry, National Chung Hsing University, Taichung, 402, Taiwan C.-I. Chang Graduate Institute of Biotechnology, National Pingtung University of Science and Technology, Pingtung, 912, Taiwan Y.-H. Tseng (&) Institute of Medical Biotechnology, Central Taiwan University of Science and Technology, Taichung, 406, Taiwan e-mail: [email protected]
Introduction Stenotrophomonas maltophilia (Sm), previously called Pseudomonas maltophilia or Xanthomonas maltophilia (Palleroni and Bradbury 1993), is an aerobic, gram-negative bacterium found in various environments, including soil, water, sediment, sewage, frozen foods, plants, and animals (Hugh 1981; Juhnke et al. 1987; Aznar and Alcaide 1992; Hauben et al. 1999; Lambert et al. 1999). As an opportunistic human pathogen, Sm has signiWcantly increased the incidence of nosocomial infection in the last decade. This
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