J. Hu, Y. Wu, L. Kang, Y. Liu, Y. Wu, D. Han, J. Wang
{"title":"recA双荧光素酶报告系统的构建及其在志贺毒素转化噬菌体诱导膳食抑制剂筛选中的应用","authors":"J. Hu, Y. Wu, L. Kang, Y. Liu, Y. Wu, D. Han, J. Wang","doi":"10.1134/S0003683824604190","DOIUrl":null,"url":null,"abstract":"<p>Shiga toxin-producing <i>Escherichia coli</i> (STEC) is an important foodborne pathogen that seriously threats the public and animal health. The major virulence of STEC is Shiga toxin (Stx) which is encoded by Stx prophage. Stx prophage induction leads to the production of Stx. Our recent research has shown that dietary functional sugars can differently regulate Shiga toxin type 2 (Stx2) prophage induction. However, high-throughput assays are required to find more dietary inhibitors for Stx prophage induction. This study aims to establish the <i>recA</i> dual-luciferase reporter system for rapidly screening the functional sugars that can inhibit Stx prophage induction. Results showed that the <i>recA</i> dual-luciferase reporter plasmid was successfully constructed as confirmed by DNA sequencing. Established <i>recA</i> dual-luciferase reporter system showed similar screening results when compared with traditional temperate phage determination method (i.e., qPCR) and suggested that L-arabinose is a dietary functional Stx prophage induction inhibitor. Supplementation with L-arabinose in culture medium reduces Stx production in STEC isolated from fecal samples of diarrhea piglets. Collectively, the <i>recA</i> dual-luciferase reporter system is a promising tool for convenient and rapid screening of dietary Stx prophage induction inhibitor, especially suitable for large-scale screening experiments.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":"60 6","pages":"1131 - 1136"},"PeriodicalIF":1.0000,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Construction of recA Dual Luciferase Reporter System and Its Application on Screening of Dietary Inhibitors for Shiga Toxin-Converting Bacteriophage Induction\",\"authors\":\"J. Hu, Y. Wu, L. Kang, Y. Liu, Y. Wu, D. Han, J. Wang\",\"doi\":\"10.1134/S0003683824604190\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Shiga toxin-producing <i>Escherichia coli</i> (STEC) is an important foodborne pathogen that seriously threats the public and animal health. The major virulence of STEC is Shiga toxin (Stx) which is encoded by Stx prophage. Stx prophage induction leads to the production of Stx. Our recent research has shown that dietary functional sugars can differently regulate Shiga toxin type 2 (Stx2) prophage induction. However, high-throughput assays are required to find more dietary inhibitors for Stx prophage induction. This study aims to establish the <i>recA</i> dual-luciferase reporter system for rapidly screening the functional sugars that can inhibit Stx prophage induction. Results showed that the <i>recA</i> dual-luciferase reporter plasmid was successfully constructed as confirmed by DNA sequencing. Established <i>recA</i> dual-luciferase reporter system showed similar screening results when compared with traditional temperate phage determination method (i.e., qPCR) and suggested that L-arabinose is a dietary functional Stx prophage induction inhibitor. Supplementation with L-arabinose in culture medium reduces Stx production in STEC isolated from fecal samples of diarrhea piglets. Collectively, the <i>recA</i> dual-luciferase reporter system is a promising tool for convenient and rapid screening of dietary Stx prophage induction inhibitor, especially suitable for large-scale screening experiments.</p>\",\"PeriodicalId\":466,\"journal\":{\"name\":\"Applied Biochemistry and Microbiology\",\"volume\":\"60 6\",\"pages\":\"1131 - 1136\"},\"PeriodicalIF\":1.0000,\"publicationDate\":\"2024-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Applied Biochemistry and Microbiology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://link.springer.com/article/10.1134/S0003683824604190\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Applied Biochemistry and Microbiology","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1134/S0003683824604190","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Construction of recA Dual Luciferase Reporter System and Its Application on Screening of Dietary Inhibitors for Shiga Toxin-Converting Bacteriophage Induction
Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogen that seriously threats the public and animal health. The major virulence of STEC is Shiga toxin (Stx) which is encoded by Stx prophage. Stx prophage induction leads to the production of Stx. Our recent research has shown that dietary functional sugars can differently regulate Shiga toxin type 2 (Stx2) prophage induction. However, high-throughput assays are required to find more dietary inhibitors for Stx prophage induction. This study aims to establish the recA dual-luciferase reporter system for rapidly screening the functional sugars that can inhibit Stx prophage induction. Results showed that the recA dual-luciferase reporter plasmid was successfully constructed as confirmed by DNA sequencing. Established recA dual-luciferase reporter system showed similar screening results when compared with traditional temperate phage determination method (i.e., qPCR) and suggested that L-arabinose is a dietary functional Stx prophage induction inhibitor. Supplementation with L-arabinose in culture medium reduces Stx production in STEC isolated from fecal samples of diarrhea piglets. Collectively, the recA dual-luciferase reporter system is a promising tool for convenient and rapid screening of dietary Stx prophage induction inhibitor, especially suitable for large-scale screening experiments.
期刊介绍:
Applied Biochemistry and Microbiology is an international peer reviewed journal that publishes original articles on biochemistry and microbiology that have or may have practical applications. The studies include: enzymes and mechanisms of enzymatic reactions, biosynthesis of low and high molecular physiologically active compounds; the studies of their structure and properties; biogenesis and pathways of their regulation; metabolism of producers of biologically active compounds, biocatalysis in organic synthesis, applied genetics of microorganisms, applied enzymology; protein and metabolic engineering, biochemical bases of phytoimmunity, applied aspects of biochemical and immunochemical analysis; biodegradation of xenobiotics; biosensors; biomedical research (without clinical studies). Along with experimental works, the journal publishes descriptions of novel research techniques and reviews on selected topics.