副溶血性弧菌:生长、遗传操作和毒力因子分析的基本技术

Current Protocols in Microbiology Pub Date : 2020-12-07 DOI:10.1002/cpmc.131

Suneeta Chimalapati, Alexander E. Lafrance, Luming Chen, Kim Orth

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引用次数: 7

摘要

副溶血性弧菌是一种革兰氏阴性的嗜盐细菌，是人类和虾类的机会致病菌。研究副溶血性弧菌感染的机制及其使用的多种毒力因素需要进行细菌培养、基因操作和毒力表型分析。详细的方案生长评估，产生突变，和表型评估包括在这篇文章。©2020 Wiley期刊有限公司基本方案1:评估副溶血性弧菌的生长替代方案1:使用平板读取器评估副溶血性弧菌的生长基本方案2:游泳/群体运动测定基本方案3:遗传操作替代方案2:自然转化基本方案4:分泌测定和质谱分析样品制备基本方案5:入侵测定(庆大霉素保护测定)基本方案6:细胞内副溶血性弧菌的免疫荧光检测基本方案7:T3SS2的细胞毒性测定

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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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来源期刊

Current Protocols in Microbiology Immunology and Microbiology-Parasitology

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期刊介绍： Current Protocols in Microbiology provides detailed, step-by-step instructions for analyzing bacteria, animal and plant viruses, fungi, protozoans and other microbes. It offers updated coverage of emerging technologies and concepts, such as biofilms, quorum sensing and quantitative PCR, as well as proteomic and genomic methods. It is the first comprehensive source of high-quality microbiology protocols that reflects and incorporates the new mandates and capabilities of this robust and rapidly evolving discipline.