Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applications
<p>Abstract</p> <p>Background</p> <p>Evolvability is an intrinsic feature of all living cells. However, newly emerging, evolved features can be undesirable when genetic circuits, designed and fabricated by rational, synthetic biological approaches, are installed in the...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2010-05-01
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| Series: | Microbial Cell Factories |
| Online Access: | http://www.microbialcellfactories.com/content/9/1/38 |
| _version_ | 1818117158467534848 |
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| author | Blattner Frederick R Pósfai János Ayaydin Ferhan Balikó Gabriella Fehér Tamás Umenhoffer Kinga Pósfai György |
| author_facet | Blattner Frederick R Pósfai János Ayaydin Ferhan Balikó Gabriella Fehér Tamás Umenhoffer Kinga Pósfai György |
| author_sort | Blattner Frederick R |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Evolvability is an intrinsic feature of all living cells. However, newly emerging, evolved features can be undesirable when genetic circuits, designed and fabricated by rational, synthetic biological approaches, are installed in the cell. Streamlined-genome <it>E. coli </it>MDS42 is free of mutation-generating IS elements, and can serve as a host with reduced evolutionary potential.</p> <p>Results</p> <p>We analyze an extreme case of toxic plasmid clone instability, and show that random host IS element hopping, causing inactivation of the toxic cloned sequences, followed by automatic selection of the fast-growing mutants, can prevent the maintenance of a clone developed for vaccine production. Analyzing the molecular details, we identify a hydrophobic protein as the toxic byproduct of the clone, and show that IS elements spontaneously landing in the cloned fragment relieve the cell from the stress by blocking transcription of the toxic gene. Bioinformatics analysis of sequence reads from early shotgun genome sequencing projects, where clone libraries were constructed and maintained in <it>E. coli</it>, suggests that such IS-mediated inactivation of ectopic genes inhibiting the growth of the <it>E. coli </it>cloning host might happen more frequently than generally anticipated, leading to genomic instability and selection of altered clones.</p> <p>Conclusions</p> <p>Delayed genetic adaptation of clean-genome, IS-free MDS42 host improves maintenance of unstable genetic constructs, and is suggested to be beneficial in both laboratory and industrial settings.</p> |
| first_indexed | 2024-12-11T04:33:58Z |
| format | Article |
| id | doaj.art-fc624064a5d444d2a7904008872a610a |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-11T04:33:58Z |
| publishDate | 2010-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-fc624064a5d444d2a7904008872a610a2022-12-22T01:20:48ZengBMCMicrobial Cell Factories1475-28592010-05-01913810.1186/1475-2859-9-38Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applicationsBlattner Frederick RPósfai JánosAyaydin FerhanBalikó GabriellaFehér TamásUmenhoffer KingaPósfai György<p>Abstract</p> <p>Background</p> <p>Evolvability is an intrinsic feature of all living cells. However, newly emerging, evolved features can be undesirable when genetic circuits, designed and fabricated by rational, synthetic biological approaches, are installed in the cell. Streamlined-genome <it>E. coli </it>MDS42 is free of mutation-generating IS elements, and can serve as a host with reduced evolutionary potential.</p> <p>Results</p> <p>We analyze an extreme case of toxic plasmid clone instability, and show that random host IS element hopping, causing inactivation of the toxic cloned sequences, followed by automatic selection of the fast-growing mutants, can prevent the maintenance of a clone developed for vaccine production. Analyzing the molecular details, we identify a hydrophobic protein as the toxic byproduct of the clone, and show that IS elements spontaneously landing in the cloned fragment relieve the cell from the stress by blocking transcription of the toxic gene. Bioinformatics analysis of sequence reads from early shotgun genome sequencing projects, where clone libraries were constructed and maintained in <it>E. coli</it>, suggests that such IS-mediated inactivation of ectopic genes inhibiting the growth of the <it>E. coli </it>cloning host might happen more frequently than generally anticipated, leading to genomic instability and selection of altered clones.</p> <p>Conclusions</p> <p>Delayed genetic adaptation of clean-genome, IS-free MDS42 host improves maintenance of unstable genetic constructs, and is suggested to be beneficial in both laboratory and industrial settings.</p>http://www.microbialcellfactories.com/content/9/1/38 |
| spellingShingle | Blattner Frederick R Pósfai János Ayaydin Ferhan Balikó Gabriella Fehér Tamás Umenhoffer Kinga Pósfai György Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applications Microbial Cell Factories |
| title | Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applications |
| title_full | Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applications |
| title_fullStr | Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applications |
| title_full_unstemmed | Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applications |
| title_short | Reduced evolvability of <it>Escherichia coli </it>MDS42, an IS-less cellular chassis for molecular and synthetic biology applications |
| title_sort | reduced evolvability of it escherichia coli it mds42 an is less cellular chassis for molecular and synthetic biology applications |
| url | http://www.microbialcellfactories.com/content/9/1/38 |
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