铁需求与铁载体产生的分析

Analysis of Iron Requirements and Siderophore Production.

作者信息

Burnside Denise M, Cianciotto Nicholas P

机构信息

Department of Microbiology and Immunology, Northwestern University Medical School, Chicago, IL, USA.

出版信息

Methods Mol Biol. 2019;1921:3-19. doi: 10.1007/978-1-4939-9048-1_1.

DOI:10.1007/978-1-4939-9048-1_1

PMID:30694482

Abstract

This chapter describes the methods for inducing, detecting, and purifying the Legionella pneumophila siderophore. The first protocol details the methods by which L. pneumophila is cultured to facilitate production of the siderophore, rhizoferrin. This chapter then describes how to purify siderophore from culture supernatants through sequential reversed-phase/weak-anion exchange chromatography and high-performance liquid chromatography. The next section describes assays which allow the detection of the iron-binding capability and the biological activity of the purified siderophore. Lastly, this chapter describes the growth of L. pneumophila in chemically defined liquid medium (CDM) containing various iron sources as a method to assess the iron requirements of L. pneumophila.

摘要

本章介绍了诱导、检测和纯化嗜肺军团菌铁载体的方法。第一个方案详细说明了培养嗜肺军团菌以促进铁载体根铁菌素产生的方法。本章接着描述了如何通过连续的反相/弱阴离子交换色谱和高效液相色谱从培养上清液中纯化铁载体。下一节介绍了用于检测纯化铁载体的铁结合能力和生物活性的分析方法。最后，本章描述了嗜肺军团菌在含有各种铁源的化学限定液体培养基（CDM）中的生长情况，以此作为评估嗜肺军团菌铁需求的一种方法。