Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer.
Storage, manipulation and delivery of DNA fragments is crucial for synthetic biology applications, subsequently allowing organisms of interest to be engineered with genes or pathways to produce desirable phenotypes such as disease or drought resistance in plants, or for synthesis of a specific chemi...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2019-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0206781 |
| _version_ | 1818934570022600704 |
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| author | Stephanie L Brumwell Michael R MacLeod Tony Huang Ryan R Cochrane Rebecca S Meaney Maryam Zamani Ola Matysiakiewicz Kaitlyn N Dan Preetam Janakirama David R Edgell Trevor C Charles Turlough M Finan Bogumil J Karas |
| author_facet | Stephanie L Brumwell Michael R MacLeod Tony Huang Ryan R Cochrane Rebecca S Meaney Maryam Zamani Ola Matysiakiewicz Kaitlyn N Dan Preetam Janakirama David R Edgell Trevor C Charles Turlough M Finan Bogumil J Karas |
| author_sort | Stephanie L Brumwell |
| collection | DOAJ |
| description | Storage, manipulation and delivery of DNA fragments is crucial for synthetic biology applications, subsequently allowing organisms of interest to be engineered with genes or pathways to produce desirable phenotypes such as disease or drought resistance in plants, or for synthesis of a specific chemical product. However, DNA with high G+C content can be unstable in many host organisms including Saccharomyces cerevisiae. Here, we report the development of Sinorhizobium meliloti, a nitrogen-fixing plant symbioticα-Proteobacterium, as a novel host that can store DNA, and mobilize DNA to E. coli, S. cerevisiae, and the eukaryotic microalgae Phaeodactylum tricornutum. To achieve this, we deleted the hsdR restriction-system in multiple reduced genome strains of S. meliloti that enable DNA transformation with up to 1.4 x 105 and 2.1 x 103 CFU μg-1 of DNA efficiency using electroporation and a newly developed polyethylene glycol transformation method, respectively. Multi-host and multi-functional shuttle vectors (MHS) were constructed and stably propagated in S. meliloti, E. coli, S. cerevisiae, and P. tricornutum. We also developed protocols and demonstrated direct transfer of these MHS vectors via conjugation from S. meliloti to E. coli, S. cerevisiae, and P. tricornutum. The development of S. meliloti as a new host for inter-kingdom DNA transfer will be invaluable for synthetic biology research and applications, including the installation and study of genes and biosynthetic pathways into organisms of interest in industry and agriculture. |
| first_indexed | 2024-12-20T05:06:22Z |
| format | Article |
| id | doaj.art-cea7a9fd87ad47e7b25e7a9700fc6982 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T05:06:22Z |
| publishDate | 2019-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-cea7a9fd87ad47e7b25e7a9700fc69822022-12-21T19:52:23ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-01146e020678110.1371/journal.pone.0206781Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer.Stephanie L BrumwellMichael R MacLeodTony HuangRyan R CochraneRebecca S MeaneyMaryam ZamaniOla MatysiakiewiczKaitlyn N DanPreetam JanakiramaDavid R EdgellTrevor C CharlesTurlough M FinanBogumil J KarasStorage, manipulation and delivery of DNA fragments is crucial for synthetic biology applications, subsequently allowing organisms of interest to be engineered with genes or pathways to produce desirable phenotypes such as disease or drought resistance in plants, or for synthesis of a specific chemical product. However, DNA with high G+C content can be unstable in many host organisms including Saccharomyces cerevisiae. Here, we report the development of Sinorhizobium meliloti, a nitrogen-fixing plant symbioticα-Proteobacterium, as a novel host that can store DNA, and mobilize DNA to E. coli, S. cerevisiae, and the eukaryotic microalgae Phaeodactylum tricornutum. To achieve this, we deleted the hsdR restriction-system in multiple reduced genome strains of S. meliloti that enable DNA transformation with up to 1.4 x 105 and 2.1 x 103 CFU μg-1 of DNA efficiency using electroporation and a newly developed polyethylene glycol transformation method, respectively. Multi-host and multi-functional shuttle vectors (MHS) were constructed and stably propagated in S. meliloti, E. coli, S. cerevisiae, and P. tricornutum. We also developed protocols and demonstrated direct transfer of these MHS vectors via conjugation from S. meliloti to E. coli, S. cerevisiae, and P. tricornutum. The development of S. meliloti as a new host for inter-kingdom DNA transfer will be invaluable for synthetic biology research and applications, including the installation and study of genes and biosynthetic pathways into organisms of interest in industry and agriculture.https://doi.org/10.1371/journal.pone.0206781 |
| spellingShingle | Stephanie L Brumwell Michael R MacLeod Tony Huang Ryan R Cochrane Rebecca S Meaney Maryam Zamani Ola Matysiakiewicz Kaitlyn N Dan Preetam Janakirama David R Edgell Trevor C Charles Turlough M Finan Bogumil J Karas Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer. PLoS ONE |
| title | Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer. |
| title_full | Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer. |
| title_fullStr | Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer. |
| title_full_unstemmed | Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer. |
| title_short | Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer. |
| title_sort | designer sinorhizobium meliloti strains and multi functional vectors enable direct inter kingdom dna transfer |
| url | https://doi.org/10.1371/journal.pone.0206781 |
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