Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene marker
<p>Abstract</p> <p>Background</p> <p>Antibiotic resistance genes are widely used for selection of recombinant bacteria, but their use risks contributing to the spread of antibiotic resistance. In particular, the practice is inappropriate for some intrinsically resistant...
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| Format: | Article |
| Language: | English |
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BMC
2008-08-01
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| Series: | BMC Biotechnology |
| Online Access: | http://www.biomedcentral.com/1472-6750/8/61 |
| _version_ | 1828463297557430272 |
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| author | Good Liam Goh Shan |
| author_facet | Good Liam Goh Shan |
| author_sort | Good Liam |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Antibiotic resistance genes are widely used for selection of recombinant bacteria, but their use risks contributing to the spread of antibiotic resistance. In particular, the practice is inappropriate for some intrinsically resistant bacteria and in vaccine production, and costly for industrial scale production. Non-antibiotic systems are available, but require mutant host strains, defined media or expensive reagents. An unexplored concept is over-expression of a host essential gene to enable selection in the presence of a chemical inhibitor of the gene product. To test this idea in <it>E. coli</it>, we used the growth essential target gene <it>fabI </it>as the plasmid-borne marker and the biocide triclosan as the selective agent.</p> <p>Results</p> <p>The new cloning vector, pFab, enabled selection by triclosan at 1 μM. Interestingly, pFab out-performed the parent pUC19-ampicillin system in cell growth, plasmid stability and plasmid yield. Also, pFab was toxic to host cells in a way that was reversed by triclosan. Therefore, pFab and triclosan are toxic when used alone but in combination they enhance growth and plasmid production through a gene-inhibitor interaction.</p> <p>Conclusion</p> <p>The <it>fabI</it>-triclosan model system provides an alternative plasmid selection method based on essential gene over-expression, without the use of antibiotic-resistance genes and conventional antibiotics.</p> |
| first_indexed | 2024-12-11T02:52:57Z |
| format | Article |
| id | doaj.art-b95b5559826140898091173078ccf60c |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-11T02:52:57Z |
| publishDate | 2008-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-b95b5559826140898091173078ccf60c2022-12-22T01:23:16ZengBMCBMC Biotechnology1472-67502008-08-01816110.1186/1472-6750-8-61Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene markerGood LiamGoh Shan<p>Abstract</p> <p>Background</p> <p>Antibiotic resistance genes are widely used for selection of recombinant bacteria, but their use risks contributing to the spread of antibiotic resistance. In particular, the practice is inappropriate for some intrinsically resistant bacteria and in vaccine production, and costly for industrial scale production. Non-antibiotic systems are available, but require mutant host strains, defined media or expensive reagents. An unexplored concept is over-expression of a host essential gene to enable selection in the presence of a chemical inhibitor of the gene product. To test this idea in <it>E. coli</it>, we used the growth essential target gene <it>fabI </it>as the plasmid-borne marker and the biocide triclosan as the selective agent.</p> <p>Results</p> <p>The new cloning vector, pFab, enabled selection by triclosan at 1 μM. Interestingly, pFab out-performed the parent pUC19-ampicillin system in cell growth, plasmid stability and plasmid yield. Also, pFab was toxic to host cells in a way that was reversed by triclosan. Therefore, pFab and triclosan are toxic when used alone but in combination they enhance growth and plasmid production through a gene-inhibitor interaction.</p> <p>Conclusion</p> <p>The <it>fabI</it>-triclosan model system provides an alternative plasmid selection method based on essential gene over-expression, without the use of antibiotic-resistance genes and conventional antibiotics.</p>http://www.biomedcentral.com/1472-6750/8/61 |
| spellingShingle | Good Liam Goh Shan Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene marker BMC Biotechnology |
| title | Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene marker |
| title_full | Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene marker |
| title_fullStr | Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene marker |
| title_full_unstemmed | Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene marker |
| title_short | Plasmid selection in <it>Escherichia coli </it>using an endogenous essential gene marker |
| title_sort | plasmid selection in it escherichia coli it using an endogenous essential gene marker |
| url | http://www.biomedcentral.com/1472-6750/8/61 |
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