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Separating Bacteria by Capsule Amount Using a Discontinuous Density Gradient.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Feltwell, Theresa 
Dorman, Matthew J 
Goulding, David A 
Short, Francesca L 

Abstract

Capsule is a key virulence factor in many bacterial species, mediating immune evasion and resistance to various physical stresses. While many methods are available to quantify and compare capsule production between different strains or mutants, there is no widely used method for sorting bacteria based on how much capsule they produce. We have developed a method to separate bacteria by capsule amount, using a discontinuous density gradient. This method is used to compare capsule amounts semi-quantitatively between cultures, to isolate mutants with altered capsule production, and to purify capsulated bacteria from complex samples. This method can also be coupled with transposon-insertion sequencing to identify genes involved in capsule regulation. Here, the method is demonstrated in detail, including how to optimize the gradient conditions for a new bacterial species or strain, and how to construct and run the density gradient.

Description

Keywords

Animals, Bacterial Capsules, Virulence Factors

Journal Title

J Vis Exp

Conference Name

Journal ISSN

1940-087X
1940-087X

Volume Title

Publisher

MyJove Corporation