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利用生物发光沙门氏菌评估构建的基于噬菌体的生物吸附剂的效率。

Use of bioluminescent Salmonella for assessing the efficiency of constructed phage-based biosorbent.

作者信息

Sun W, Brovko L, Griffiths M

机构信息

Department of Food Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1.

出版信息

J Ind Microbiol Biotechnol. 2001 Aug;27(2):126-8. doi: 10.1038/sj.jim.7000198.

Abstract

A bacteriophage-based biosorbent for Salmonella enteritidis was constructed, and bacterial bioluminescence was used for assessment of the efficiency of cell capture. A strain of S. enteritidis with bioluminescent phenotype was constructed by transformation with plasmid pT7 carrying the entire lux operon from Photorhabdus luminescens. The relation between relative light output (RLU) and colony-forming units (CFU/ml) of the bioluminescent strain was established. The bacteriophage specific to S. enteritidis was biotinylated, and the biotinylation procedure was optimized based on the maximum retention of phage infectivity. The biotinylated phages were then coated onto streptavidin-labeled magnetic beads, and were used to capture the bioluminescent S. enteritidis cells. Our preliminary results showed that the number of cells captured by constructed biosorbent was five times higher than that of the control, magnetic beads coated with nonbiotinylated phage, indicating the capture is specific.

摘要

构建了一种基于噬菌体的肠炎沙门氏菌生物吸附剂,并利用细菌生物发光来评估细胞捕获效率。通过用携带来自发光杆菌属完整lux操纵子的质粒pT7转化,构建了具有生物发光表型的肠炎沙门氏菌菌株。建立了生物发光菌株的相对光输出(RLU)与菌落形成单位(CFU/ml)之间的关系。对肠炎沙门氏菌特异性噬菌体进行生物素化,并基于噬菌体感染性的最大保留对生物素化程序进行了优化。然后将生物素化的噬菌体包被在链霉亲和素标记的磁珠上,用于捕获生物发光的肠炎沙门氏菌细胞。我们的初步结果表明,构建的生物吸附剂捕获的细胞数量比对照(包被未生物素化噬菌体的磁珠)高五倍,表明捕获具有特异性。

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