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培养并鉴定感染水华蓝藻铜绿微囊藻的 MaMV-DC 噬藻体。

Cultivation and characterization of the MaMV-DC cyanophage that infects bloom-forming cyanobacterium Microcystis aeruginosa.

机构信息

State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China.

出版信息

Virol Sin. 2013 Oct;28(5):266-71. doi: 10.1007/s12250-013-3340-7. Epub 2013 Aug 26.

DOI:10.1007/s12250-013-3340-7
PMID:23990146
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8208409/
Abstract

The MaMV-DC cyanophage, which infects the bloom-forming cyanobacterium Microcystis aeruginosa, was isolated from Lake Dianchi, Kunming, China. Twenty-one cyanobacterial strains were used to detect the host range of MaMV-DC. Microcystic aeruginosa FACHB-524 and plaque purification were used to isolate individual cyanophages, and culturing MaMV-DC with cyanobacteria allowed us to prepare purified cyanophages for further analysis. Electron microscopy demonstrated that the negatively stained viral particles are tadpole-shaped with an icosahedral head approximately 70 nm in diameter and a contractile tail approximately 160 nm in length. Using one-step growth experiments, the latent period and burst size of MaMV-DC were estimated to be 24-48 hours and approximately 80 infectious units per cell, respectively. Restriction endonuclease digestion and agarose gel electrophoresis were performed using purified MaMV-DC genomic DNA, and the genome size was estimated to be approximately 160 kb. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed four major structural proteins. These results support the growing interest in using freshwater cyanophages to control bloom-forming cyanobacterium.

摘要

感染水华蓝藻铜绿微囊藻的 MaMV-DC 噬藻体,从中国昆明滇池中分离得到。用 21 株蓝藻菌株检测 MaMV-DC 的宿主范围。采用微囊藻 FACHB-524 和噬菌斑纯化的方法分离出单个噬藻体,用噬藻体与蓝藻共培养的方法,我们制备了纯化的噬藻体,以进行进一步分析。电子显微镜显示,负染的病毒颗粒呈蝌蚪状,具有直径约 70nm 的二十面体头部和长约 160nm 的可收缩尾部。通过一步生长实验,估计 MaMV-DC 的潜伏期和爆发量分别为 24-48 小时和每个细胞约 80 个感染单位。用纯化的 MaMV-DC 基因组 DNA 进行限制性内切酶消化和琼脂糖凝胶电泳,基因组大小估计约为 160kb。十二烷基硫酸钠聚丙烯酰胺凝胶电泳 (SDS-PAGE) 分析显示有 4 种主要的结构蛋白。这些结果支持了利用淡水噬藻体来控制水华蓝藻的日益增长的兴趣。

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