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Wiley, Current Protocols in Microbiology, 1(36), 2015

DOI: 10.1002/9780471729259.mc01e03s36

Wiley, Current Protocols in Microbiology, 1(19), 2010

DOI: 10.1002/9780471729259.mc01e03s19

Wiley, Current Protocols in Molecular Biology, 1(106), 2014

DOI: 10.1002/0471142727.mb0716s106

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Genome-Wide Fitness and Genetic Interactions Determined by Tn-seq, a High-Throughput Massively Parallel Sequencing Method for Microorganisms

Journal article published in 2010 by Tim van Opijnen ORCID, David W. Lazinski, Andrew Camilli
This paper is available in a repository.
This paper is available in a repository.

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Abstract

The lagging annotation of bacterial genomes and the inherent genetic complexity of many phenotypes is hindering the discovery of new drug targets and the development of new antimicrobials and vaccines. Here we present the method Tn-seq, with which it has become possible to quantitatively determine fitness for most genes in a microorganism and to screen for quantitative genetic interactions on a genome-wide scale and in a high-throughput fashion. Tn-seq can thus direct studies in the annotation of genes and untangle complex phenotypes. The method is based on the construction of a saturated Mariner transposon insertion library. After library selection, changes in frequency of each insertion mutant are determined by sequencing of the flanking regions en masse. These changes are used to calculate each mutant’s fitness. The method has been developed for the Gram-positive bacterium Streptococcus pneumoniae, a causative agent of pneumonia and meningitis; however, due to the wide activity of the Mariner transposon, Tn-seq can be applied to many different microbial species.