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首页> 外文期刊>Applied Microbiology >Cell Surface Display of Poliovirus Receptor on Escherichia coli, a Novel Method for Concentrating Viral Particles in Water
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Cell Surface Display of Poliovirus Receptor on Escherichia coli, a Novel Method for Concentrating Viral Particles in Water

机译：脊髓灰质炎病毒受体在大肠杆菌上的细胞表面展示，一种浓缩水中病毒颗粒的新方法

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摘要

The lack of efficient methods for concentrating viruses in water samples leads to underreporting of viral contamination in source water. A novel strategy for viral concentration was developed using the expression of target virus receptors on bacterial cells. Poliovirus type 1, the most studied enterovirus, was used as a surrogate for enteric viruses. The human poliovirus receptor ( hPVR ) gene was expressed on the surface of Escherichia coli cells by using the ice nucleation protein ( INP ) gene. The hPVR gene was ligated to the 3′ end of the INP gene after the removal of the stop codon. The resulting open reading frame (ORF) was used for the projection of hPVR onto the outer membrane of E. coli . Gene expression was tested by SDS-PAGE, Western blot, and dot blot analyses, and virion capture ability was confirmed by transmission electron microscopy. The application of engineered E. coli cells for capturing viruses in 1-liter samples of source and drinking water resulted in 75 to 99% procedural recovery efficiency. Cell surface display of viral receptors on bacterial cells opens a new prospect for an efficient and inexpensive alternative tool for capturing and concentrating waterborne viruses in water samples.
机译：缺乏有效的方法来浓缩水样中的病毒会导致对源水中病毒污染的报告不足。利用靶病毒受体在细菌细胞上的表达，开发了一种新的病毒浓缩策略。脊髓灰质炎病毒1型（研究最深入的肠道病毒）被用作肠道病毒的替代品。利用冰核蛋白（INP）基因在大肠杆菌细胞表面表达人类脊髓灰质炎病毒受体（hPVR）基因。去除终止密码子后，将hPVR基因连接到INP基因的3'末端。所得的开放阅读框（ORF）用于将hPVR投射到大肠杆菌的外膜上。通过SDS-PAGE，蛋白质印迹和斑点印迹分析测试基因表达，并通过透射电子显微镜确认病毒体捕获能力。应用工程化的大肠杆菌细胞捕获1升源水和饮用水样品中的病毒可以使程序回收率达到75％到99％。细菌细胞上病毒受体在细胞表面的展示，为捕获和浓缩水样中的水传播病毒提供了一种有效而廉价的替代工具的新前景。

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