2017
DOI: 10.3389/fmicb.2017.01842
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Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans

Abstract: Aggregatibacter actinomycetemcomitans is the primary etiological agent of aggressive periodontal disease. Identification of novel virulence factors at the genome-wide level is hindered by lack of efficient genetic tools to perform mutagenesis in this organism. The Himar1 mariner transposon is known to yield a random distribution of insertions in an organism’s genome with requirement for only a TA dinucleotide target and is independent of host-specific factors. However, the utility of this system in A. actinomy… Show more

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Cited by 11 publications
(6 citation statements)
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“…Transposon sequencing (Tn-seq) is a powerful forward genetic screening tool that has been successfully employed for genetic fitness studies of a number of pathogens ( 34 ) and pathobiont microbiome species ( 25 , 35 ). Given the high level of natural competence observed in P. micra , we were curious to determine the feasibility of implementing in vitro transposon mutagenesis using recombinant MarC9 transposase and the mini- mariner transposon commonly used in Tn-seq studies ( Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Transposon sequencing (Tn-seq) is a powerful forward genetic screening tool that has been successfully employed for genetic fitness studies of a number of pathogens ( 34 ) and pathobiont microbiome species ( 25 , 35 ). Given the high level of natural competence observed in P. micra , we were curious to determine the feasibility of implementing in vitro transposon mutagenesis using recombinant MarC9 transposase and the mini- mariner transposon commonly used in Tn-seq studies ( Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Before the experiments, it is also important to remember that some transposons have specific insertion target sites that increase the abundance of mutants at certain loci in the genome 30 . Mariner transposons are known to target AT sites for insertion 31 , and Tn5 transposons have a GC bias 32 , 33 . Including steps during bioinformatics analysis to recognize hotspots for transposon insertions will help in assessing any distribution bias.…”
Section: Discussionmentioning
confidence: 99%
“…Transposon sequencing (Tn-seq) is a powerful forward genetic screening tool that has been successfully employed for genetic fitness studies of a number of pathogens [33] and pathobiont microbiome species [25,34]. Given the high level of natural competence observed in P. micra, we were curious to determine the feasibility of implementing in vitro transposon mutagenesis using recombinant MarC9 transposase and the mini-Mariner transposon commonly used in Tn-seq studies (Figure 7A-B) [25,33,35].…”
Section: Development Of In Vitro Transposon Mutagenesis For Tn-seq Studiesmentioning
confidence: 99%