115
5.4 High Throughput Screening Approaches
5.4.1 Transposon Mutant Library Screening
Transposons are genetic elements that can jump from one genomic location to
another, which can profoundly influence gene expression. Transposon mutagenesis
is a powerful approach to generate randomized gene mutations among bacterial
genomes. This technique has been widely used to identify individual genes involved
Table 5.2 High throughput screening approaches in persisters studies
Organisms
Tools
Library
scale
Number
of genes Antibiotic
Reference
Transposon mutant screening
P. aeruginosa
PA14
pTnMod-OGm
plasposon
5000
9
Ofloxacin
De Groote et al.
(2009)
E. coli
mini-Tn10
11,748
1
Ampicillin
Li and Zhang
(2007)
P. aeruginosa
PAO1
ISphoA/hah
5000
1
Carbenicillin
Manuel et al.
(2010)
S. aureus
USA500
pBTn
9120 13
Levofloxacin
Wang et al.
(2015)
M. tuberculosis IS1096 based
pCG113 plasmid
576
1
Isoniazidtreated mice
Camacho et al.
(1999) and Dhar
and McKinney
(2010)
Mycobacterium
fortuitum
TnphoA
125
1
Mice infection
model
Parti et al. (2008)
M. bovis BCG
Tn5370
3500
1
Deficient in
cord formation
Glickman et al.
(2000)
E. coli K12
Mini-Tn10
5000
2
Kanamycin
Hu and Coates
(2005)
S. aureus
Newman
pTM239pTM244
2
Ciprofloxacin
Wang et al.
(2018)
Transposon sequencing
E. coli MG1655 mini-Tn10
200,000
Gentamicin
Shan et al. (2015)
Uropathogenic
E. coli
EZ-Tn5
360,000
Ampicillin
Molina-Quiroz
et al. (2016)
P. aeruginosa
PAO1
pIT2-Tn5
100,000
Ciprofloxacin
Cameron et al.
(2018)
Deletion mutant library screening
E. coli K-12
Knockout
3985
2
Ampicillin
Ma et al. (2010)
E. coli K-12
Knockout
3985 37
Ofloxacin
Cui et al. (2018)
Overexpression library screening
E. coli K-12
Overexpression
4500
2
Ampicillin
Spoering et al.
(2006)
5 Molecular and Systems Biology Approaches for Analyzing Drug-Tolerant Bacterial…