Imperial College London

DrJanineBosse

Faculty of MedicineDepartment of Infectious Disease

Senior Research Fellow
 
 
 
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Contact

 

+44 (0)20 7594 1803j.bosse

 
 
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Location

 

234Wright Fleming WingSt Mary's Campus

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Summary

 

Publications

Citation

BibTex format

@article{Bossé:2006:10.2174/187152606778249917,
author = {Bossé, JT and Zhou, L and Kroll, JS and Langford, PR},
doi = {10.2174/187152606778249917},
journal = {Infect Disord Drug Targets},
pages = {241--262},
title = {High-throughput identification of conditionally essential genes in bacteria: from STM to TSM.},
url = {http://dx.doi.org/10.2174/187152606778249917},
volume = {6},
year = {2006}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - Signature-tagged mutagenesis (STM) provided the first widely applicable high-throughput method for detecting conditionally essential genes in bacteria by using negative selection to screen large pools of transposon (Tn) mutants. STM requires no prior knowledge of the bacterium's genome sequence, and has been used to study a large number of Gram-positive and Gram-negative species, greatly expanding the repertoires of known virulence factors for these organisms. Originally, hybridization of radiolabelled probes to colony or dot blots was used to detect differences in populations of tagged mutants before and after growth under a selective condition. Modifications of the tag detection method involving polymerase chain reaction (PCR) amplification and visualisation by gel electrophoresis have been developed and can be automated through the use of robotics. Genetic footprinting is another negative selection technique that uses PCR amplification to detect loss of mutants from a pool. Unlike PCR-STM, this technique allows direct amplification of Tn-flanking sequences. However, it requires the bacterium's whole genome sequence in order to design specific primers for every gene of interest. More recently, a number of techniques have been described that combine the negative-selection principle of STM and genetic footprinting with the genome-wide screening power of DNA microarrays. These techniques, although also requiring whole genome sequences, use either a form of linker-mediated or semi-random PCR to amplify and label Tn-flanking regions for hybridization to microarrays. The superior sensitivity microarray detection allows greater numbers of mutants to be screened per pool, as well as determination of the coverage/distribution of insertions in the library prior to screening, two significant advantages over STM.
AU - Bossé,JT
AU - Zhou,L
AU - Kroll,JS
AU - Langford,PR
DO - 10.2174/187152606778249917
EP - 262
PY - 2006///
SN - 1871-5265
SP - 241
TI - High-throughput identification of conditionally essential genes in bacteria: from STM to TSM.
T2 - Infect Disord Drug Targets
UR - http://dx.doi.org/10.2174/187152606778249917
UR - https://www.ncbi.nlm.nih.gov/pubmed/16918485
VL - 6
ER -