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副溶血性弧菌：生长、遗传操作和毒力因子分析的基本技术。

Vibrio parahaemolyticus: Basic Techniques for Growth, Genetic Manipulation, and Analysis of Virulence Factors.

机构信息

Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas.

Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, Texas.

出版信息

Curr Protoc Microbiol. 2020 Dec;59(1):e131. doi: 10.1002/cpmc.131.

DOI:10.1002/cpmc.131
PMID:33285040
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7727304/
Abstract

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.

摘要

副溶血性弧菌是一种革兰氏阴性、嗜盐细菌，也是人类和虾类的机会性病原体。研究副溶血性弧菌感染的机制和它所采用的多种毒力因子，需要进行细菌培养、遗传操作和毒力表型分析。本文包括了生长评估、突变体生成和表型评估的详细方案。 2020 年威立出版社股份有限公司。 基本方案 1：副溶血性弧菌生长评估 可选方案 1：使用平板读数器评估副溶血性弧菌的生长 基本方案 2：泳动/扩散运动测定法 基本方案 3：遗传操作 可选方案 2：自然转化 基本方案 4：分泌测定法和用于质谱分析的样品制备 基本方案 5：侵袭测定法（庆大霉素保护测定法） 基本方案 6：胞内副溶血性弧菌的免疫荧光检测 基本方案 7：T3SS2 的细胞毒性测定法

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/316963c2b420/nihms-1644861-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/7de2c335d8ea/nihms-1644861-f0001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/316963c2b420/nihms-1644861-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/7de2c335d8ea/nihms-1644861-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/291ec2f35b8a/nihms-1644861-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/051dca74d0a3/nihms-1644861-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/ad29180a4759/nihms-1644861-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/ab0e562969d2/nihms-1644861-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc68/7727304/316963c2b420/nihms-1644861-f0006.jpg

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