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针对一株对抗生素耐药的临床大肠杆菌分离株具有裂解活性的噬菌体 fEg-Eco19 的生物学和分子特征分析。

Biological and molecular characterization of fEg-Eco19, a lytic bacteriophage active against an antibiotic-resistant clinical Escherichia coli isolate.

机构信息

Department of Bacteriology and Immunology, Medicum, Human Microbiome Research Program, Faculty of Medicine, University of Helsinki, 00014 UH, Helsinki, Finland.

Department of Botany and Microbiology, Faculty of Science, Damietta University, New Damietta, 34517, Egypt.

出版信息

Arch Virol. 2022 May;167(5):1333-1341. doi: 10.1007/s00705-022-05426-6. Epub 2022 Apr 10.

Abstract

Characterization of bacteriophages facilitates better understanding of their biology, host specificity, genomic diversity, and adaptation to their bacterial hosts. This, in turn, is important for the exploitation of phages for therapeutic purposes, as the use of uncharacterized phages may lead to treatment failure. The present study describes the isolation and characterization of a bacteriophage effective against the important clinical pathogen Escherichia coli, which shows increasing accumulation of antibiotic resistance. Phage fEg-Eco19, which is specific for a clinical E. coli strain, was isolated from an Egyptian sewage sample. Phage fEg-Eco19 formed clear, sharp-edged, round plaques. Electron microscopy showed that the isolated phage is tailed and therefore belongs to the order Caudovirales, and morphologically, it resembles siphoviruses. The diameter of the icosahedral head of fEg-Eco19 is 68 ± 2 nm, and the non-contractile tail length and diameter are 118 ± 0.2 and 13 ± 0.6 nm, respectively. The host range of the phage was found to be narrow, as it infected only two out of 137 clinical E. coli strains tested. The phage genome is 45,805 bp in length with a GC content of 50.3% and contains 76 predicted genes. Comparison of predicted and experimental restriction digestion patterns allowed rough mapping of the physical ends of the phage genome, which was confirmed using the PhageTerm tool. Annotation of the predicted genes revealed gene products belonging to several functional groups, including regulatory proteins, DNA packaging and phage structural proteins, host lysis proteins, and proteins involved in DNA/RNA metabolism and replication.

摘要

噬菌体的特性分析有助于更好地理解其生物学特性、宿主特异性、基因组多样性和对宿主细菌的适应性。这对于利用噬菌体进行治疗目的至关重要,因为使用未经特性分析的噬菌体可能导致治疗失败。本研究描述了一种针对重要临床病原体大肠杆菌的噬菌体的分离和特性分析,该菌对抗生素的耐药性逐渐增加。噬菌体 fEg-Eco19 是从埃及污水样本中分离出来的,专门针对一种临床大肠杆菌菌株。噬菌体 fEg-Eco19 形成清晰、边缘锐利、圆形的噬菌斑。电子显微镜显示,分离出的噬菌体有尾巴,因此属于尾病毒目,形态上类似于长尾噬菌体。fEg-Eco19 的二十面体头部直径为 68 ± 2nm,非收缩性尾部长度和直径分别为 118 ± 0.2nm 和 13 ± 0.6nm。噬菌体的宿主范围较窄,因为它仅感染了测试的 137 株临床大肠杆菌菌株中的两株。噬菌体基因组长 45805bp,GC 含量为 50.3%,包含 76 个预测基因。预测和实验限制酶消化图谱的比较允许对噬菌体基因组的物理末端进行大致映射,使用 PhageTerm 工具进行了确认。预测基因的注释揭示了属于几个功能组的基因产物,包括调控蛋白、DNA 包装和噬菌体结构蛋白、宿主裂解蛋白以及参与 DNA/RNA 代谢和复制的蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4249/9038960/87ce5569dcc4/705_2022_5426_Fig1_HTML.jpg

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