Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans
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...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01842/full |
| _version_ | 1818549962141597696 |
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| author | Qinfeng Ding Kai Soo Tan |
| author_facet | Qinfeng Ding Kai Soo Tan |
| author_sort | Qinfeng Ding |
| collection | DOAJ |
| description | 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. actinomycetemcomitans is unknown. In this study, we found that Himar1 transposon mutagenesis occurs at a high frequency (×10-4), and can be universally applied to wild-type A. actinomycetemcomitans strains of serotypes a, b, and c. The Himar1 transposon inserts were stably inherited in A. actinomycetemcomitans transconjugants in the absence of antibiotics. A library of 16,000 mutant colonies of A. actinomycetemcomitans was screened for reduced biofilm formation. Mutants with transposon inserts in genes encoding pilus, putative ion transporters, multidrug resistant proteins, transcription regulators and enzymes involved in the synthesis of extracellular polymeric substance, bacterial metabolism and stress response were discovered in this screen. Our results demonstrated the utility of the Himar1 mutagenesis system as a novel genetic tool for functional genomic analysis in A. actinomycetemcomitans. |
| first_indexed | 2024-12-12T08:40:06Z |
| format | Article |
| id | doaj.art-d6a4411a2801446bb98dee89497fcd92 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-12T08:40:06Z |
| publishDate | 2017-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-d6a4411a2801446bb98dee89497fcd922022-12-22T00:30:49ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-09-01810.3389/fmicb.2017.01842286366Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitansQinfeng DingKai Soo TanAggregatibacter 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. actinomycetemcomitans is unknown. In this study, we found that Himar1 transposon mutagenesis occurs at a high frequency (×10-4), and can be universally applied to wild-type A. actinomycetemcomitans strains of serotypes a, b, and c. The Himar1 transposon inserts were stably inherited in A. actinomycetemcomitans transconjugants in the absence of antibiotics. A library of 16,000 mutant colonies of A. actinomycetemcomitans was screened for reduced biofilm formation. Mutants with transposon inserts in genes encoding pilus, putative ion transporters, multidrug resistant proteins, transcription regulators and enzymes involved in the synthesis of extracellular polymeric substance, bacterial metabolism and stress response were discovered in this screen. Our results demonstrated the utility of the Himar1 mutagenesis system as a novel genetic tool for functional genomic analysis in A. actinomycetemcomitans.http://journal.frontiersin.org/article/10.3389/fmicb.2017.01842/fullmutagenesisconjugationbiofilmadherenceDNA transposable elementsperiodontal disease |
| spellingShingle | Qinfeng Ding Kai Soo Tan Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans Frontiers in Microbiology mutagenesis conjugation biofilm adherence DNA transposable elements periodontal disease |
| title | Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans |
| title_full | Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans |
| title_fullStr | Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans |
| title_full_unstemmed | Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans |
| title_short | Himar1 Transposon for Efficient Random Mutagenesis in Aggregatibacter actinomycetemcomitans |
| title_sort | himar1 transposon for efficient random mutagenesis in aggregatibacter actinomycetemcomitans |
| topic | mutagenesis conjugation biofilm adherence DNA transposable elements periodontal disease |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01842/full |
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