Development of a method for eva l uat i ng the host range of bacteriophages

开发评估噬菌体宿主范围的方法

• 批准号: 23657025
• 负责人: IMACHI Hiroyuki
• 金额: $ 2.5万
• 依托单位: Japan Agency for Marine-Earth Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-23657025/
• 关键词: ファージ; 宿主域; 16S rRNA 遺伝子; FISH 法; 16S rRNA遺伝子; FISH法

We developed a novel phage-host range evaluation method using EdU (5-ethynyl-2’-deoxyuridine) labeled phages, which consists of the following four steps. (i) Fluorescently labeled phages are added to a microbial consortium, and host cells are infected and fluorescently labeled. (ii) Fluorescent cells are sorted by FACS. (iii) 16S rRNA gene sequences retrieved from sorted cells are analyzed, and specific oligonucleotide probes for FISH are designed. (iv) Cells labeled with both fluorescently labeled phage and FISH probe are identified as host cells.

我们建立了一种利用EdU（5-乙基-2 ' -脱氧尿苷）标记噬菌体的噬菌体-宿主范围评估方法，该方法包括以下四个步骤。(i)将荧光标记的噬菌体加入微生物联合体，感染宿主细胞并进行荧光标记。（ii）荧光细胞用流式细胞仪进行分类。（iii）分析从分选细胞中检索到的16S rRNA基因序列，设计FISH特异性寡核苷酸探针。（iv）用荧光标记的噬菌体和FISH探针标记的细胞被鉴定为宿主细胞。

项目成果

過小評価されているファージの宿主域

未被充分认识的噬菌体宿主范围

• 发表时间: 2011
• 期刊: 生物工学会誌バイオミディア
• 作者: Yuichi Mizutani;Naoki Tomita;Yasuaki Niizuma;Ken Yoda;井町寛之
• 通讯作者: 井町寛之

A method for evaluating the host range of bacteriophages using phages fluorescently labeled with 5-ethynyl-2′-deoxyuridine (EdU)

使用 5-乙炔基-2-脱氧尿苷 (EdU) 荧光标记的噬菌体评估噬菌体宿主范围的方法

• DOI: 10.1007/s00253-012-4174-1
• 发表时间: 2012
• 期刊: Applied Microbiology and Biotechnology
• 影响因子: 5
• 作者: 小原良和;高橋恒二;神納健太郎;山中一司;Yusuke Takahashi; Takato Mitsudome; Tomoo Mizugaki; Koichiro Jitsukawa; Kiyotomi Kaneda;御纏真美子，平山道世，高山大鑑，岩瀬顕秀，工藤昭彦，根岸雄一;Sayaka Ohno
• 通讯作者: Sayaka Ohno