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MarTrack: A versatile toolbox of mariner transposon derivatives used for functional genetic analysis of bacterial genomes.

Abstract：

:The mariner transposon family of Himar1 has been widely used for the random mutagenesis of bacteria to generate single insertions into the chromosome. Here, a versatile toolbox of mariner transposon derivatives was generated and applied to the functional genomics investigation of fish pathogen Edwardsiella piscicida. In this study, we combined the merits of the random mutagenesis of mariner transposon and common efficient reporter marker genes or regulatory elements, mcherry, gfp, luxAB, lacZ, sacBR, and PBAD and antibiotic resistance cassettes to construct a series of derivative transposon vectors, pMmch, pMKGR, pMCGR, pMXKGR, pMLKGR, pMSGR, and pMPR, based on the initial transposon pMar2xT7. The function and effectiveness of the modified transposons were verified by introducing them into E. piscicida EIB202. Based on the toolbox, a transposon insertion mutant library containing approximately 3.0 × 105 separated mutants was constructed to explore the upstream regulators of esrB, the master regulator of the type III and type VI secretion systems (T3/T6SS) in E. piscicida. Following analysis by Con-ARTIST, ETAE_2184 (renamed as EsrR) was screened out and identified as a novel regulator mediating T3SS expression. In addition, the esrR mutants displayed critical virulence attenuation. Due to the broad-range host compatibility of mariner transposons, the newly built transposon toolbox can be broadly applied for functional genomics studies in various bacteria.

journal_name

Microbiol Res

journal_title

Microbiological research

authors

Wei L,Qiao H,Liu B,Yin K,Liu Q,Zhang Y,Ma Y,Wang Q

doi

10.1016/j.micres.2018.11.008

subject

Has Abstract

pub_date

2019-02-01 00:00:00

pages

84-93

eissn

0944-5013

issn

1618-0623

pii

S0944-5013(18)31221-7

journal_volume

219

pub_type

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