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{"title":"副溶血性弧菌:生长、遗传操作和毒力因子分析的基本技术","authors":"Suneeta Chimalapati, Alexander E. Lafrance, Luming Chen, Kim Orth","doi":"10.1002/cpmc.131","DOIUrl":null,"url":null,"abstract":"<p><i>Vibrio parahaemolyticus</i> is a Gram-negative, halophilic bacterium and opportunistic pathogen of humans and shrimp. Investigating the mechanisms of <i>V. parahaemolyticus</i> infection and the multifarious virulence factors it employs requires procedures for bacterial culture, genetic manipulation, and analysis of virulence phenotypes. Detailed protocols for growth assessment, generation of mutants, and phenotype assessment are included in this article. © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Assessment of growth of <i>V. parahaemolyticus</i></p><p><b>Alternate Protocol 1</b>: Assessment of growth of <i>V. parahaemolyticus</i> using a plate reader</p><p><b>Basic Protocol 2</b>: Swimming/swarming motility assay</p><p><b>Basic Protocol 3</b>: Genetic manipulation</p><p><b>Alternate Protocol 2</b>: Natural transformation</p><p><b>Basic Protocol 4</b>: Secretion assay and sample preparation for mass spectrometry analysis</p><p><b>Basic Protocol 5</b>: Invasion assay (gentamicin protection assay)</p><p><b>Basic Protocol 6</b>: Immunofluorescence detection of intracellular <i>V. parahaemolyticus</i></p><p><b>Basic Protocol 7</b>: Cytotoxicity assay for T3SS2</p>","PeriodicalId":39967,"journal":{"name":"Current Protocols in Microbiology","volume":"59 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmc.131","citationCount":"7","resultStr":"{\"title\":\"Vibrio parahaemolyticus: Basic Techniques for Growth, Genetic Manipulation, and Analysis of Virulence Factors\",\"authors\":\"Suneeta Chimalapati, Alexander E. Lafrance, Luming Chen, Kim Orth\",\"doi\":\"10.1002/cpmc.131\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><i>Vibrio parahaemolyticus</i> is a Gram-negative, halophilic bacterium and opportunistic pathogen of humans and shrimp. Investigating the mechanisms of <i>V. parahaemolyticus</i> infection and the multifarious virulence factors it employs requires procedures for bacterial culture, genetic manipulation, and analysis of virulence phenotypes. Detailed protocols for growth assessment, generation of mutants, and phenotype assessment are included in this article. © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Assessment of growth of <i>V. parahaemolyticus</i></p><p><b>Alternate Protocol 1</b>: Assessment of growth of <i>V. parahaemolyticus</i> using a plate reader</p><p><b>Basic Protocol 2</b>: Swimming/swarming motility assay</p><p><b>Basic Protocol 3</b>: Genetic manipulation</p><p><b>Alternate Protocol 2</b>: Natural transformation</p><p><b>Basic Protocol 4</b>: Secretion assay and sample preparation for mass spectrometry analysis</p><p><b>Basic Protocol 5</b>: Invasion assay (gentamicin protection assay)</p><p><b>Basic Protocol 6</b>: Immunofluorescence detection of intracellular <i>V. parahaemolyticus</i></p><p><b>Basic Protocol 7</b>: Cytotoxicity assay for T3SS2</p>\",\"PeriodicalId\":39967,\"journal\":{\"name\":\"Current Protocols in Microbiology\",\"volume\":\"59 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-12-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/cpmc.131\",\"citationCount\":\"7\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Protocols in Microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.131\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.131","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 7
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Vibrio parahaemolyticus: Basic Techniques for Growth, Genetic Manipulation, and Analysis of Virulence Factors
Vibrio parahaemolyticus is a Gram-negative, halophilic bacterium and opportunistic pathogen of humans and shrimp. Investigating the mechanisms of V. parahaemolyticus infection and the multifarious virulence factors it employs requires procedures for bacterial culture, genetic manipulation, and analysis of virulence phenotypes. Detailed protocols for growth assessment, generation of mutants, and phenotype assessment are included in this article. © 2020 Wiley Periodicals LLC.
Basic Protocol 1 : Assessment of growth of V. parahaemolyticus
Alternate Protocol 1 : Assessment of growth of V. parahaemolyticus using a plate reader
Basic Protocol 2 : Swimming/swarming motility assay
Basic Protocol 3 : Genetic manipulation
Alternate Protocol 2 : Natural transformation
Basic Protocol 4 : Secretion assay and sample preparation for mass spectrometry analysis
Basic Protocol 5 : Invasion assay (gentamicin protection assay)
Basic Protocol 6 : Immunofluorescence detection of intracellular V. parahaemolyticus
Basic Protocol 7 : Cytotoxicity assay for T3SS2
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