Detection of bacteria using foreign DNA: the development of a bacteriophage reagent for Salmonella

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• 作者：Kuhn ; Jonathan ; Suissa ; Mordechai ; Wyse ; Joseph ; Cohen ; Ilana ; Weiser ; Irit ; Reznick ; Sarah ; et. al.
• 关键词：Salmonella detection ; Bacteriophage reagent ; Felix 01
• 刊名：International Journal of Food Microbiology
• 出版年：2002
• 出版时间：April 5, 2002
• 年：2002
• 卷：74
• 期：3
• 页码：229-238
• 全文大小：130 K

文摘

A phage-based reagent was developed for the detection of Salmonella in food samples. The parental phage was Felix 01, which lyses practically all Salmonella. Using data obtained about the molecular biology of the phage, a recombinant phage that carried the bacterial genes specifying luciferase was produced. The method involved the isolation of amber nonsense mutations and subsequent crosses to render doubly mutant phage with a very low reversion rate on strains lacking an amber suppressor. A plasmid was constructed that contained a segment of Felix 01 DNA with two adjacent genes, one dispensable and the other essential, and their flanking sequences. Recombinant DNA technology was used to remove the two genes and the luxA and luxB genes for luciferase, and a gene specifying a tRNA that recognizes amber codons (supF