Comprehensive Essentiality Analysis of the Mycobacterium Tuberculosis Genome Via Saturating Transposon Mutagenesis

Overview

Journal mBio

Publisher American Society for Microbiology

Specialty Microbiology

Date 2017 Jan 19

PMID 28096490

Citations 341

Authors

Michael A DeJesus

Elias R Gerrick

Weizhen Xu

Sae Woong Park

Jarukit E Long

Cara C Boutte

Eric J Rubin

Dirk Schnappinger

Sabine Ehrt

Sarah M Fortune

Christopher M Sassetti

Thomas R Ioerger

Affiliations

Soon will be listed here.

Abstract

Importance: Sequencing of transposon-insertion mutant libraries has become a widely used tool for probing the functions of genes under various conditions. The Himar1 transposon is generally believed to insert with equal probabilities at all TA dinucleotides, and therefore its absence in a mutant library is taken to indicate biological selection against the corresponding mutant. Through sequencing of a saturated Himar1 library, we found evidence that TA dinucleotides are not equally permissive for insertion. The insertion bias was observed in multiple prokaryotes and influences the statistical interpretation of transposon insertion (TnSeq) data and characterization of essential genomic regions. Using these insights, we analyzed a fully saturated TnSeq library for M. tuberculosis, enabling us to generate a comprehensive catalog of in vitro essentiality, including ORFs smaller than those found in any previous study, small (noncoding) RNAs (sRNAs), promoters, and other genomic features.

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