Streamlined assembly of cloning and genome editing vectors for genus Clostridium
Summary: Reported herein is a new set of vectors designed to streamline molecular cloning and genome editing by exploiting modern cloning methods. The new vectors build on the existing pMTL8000 vectors that have been a staple of Clostridium research for more than a decade. The introduction of two pa...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-08-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004223015614 |
| _version_ | 1827868328534736896 |
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| author | Tom S. Bailey Philip Hittmeyer Yanchao Zhang Aleksandra M. Kubiak |
| author_facet | Tom S. Bailey Philip Hittmeyer Yanchao Zhang Aleksandra M. Kubiak |
| author_sort | Tom S. Bailey |
| collection | DOAJ |
| description | Summary: Reported herein is a new set of vectors designed to streamline molecular cloning and genome editing by exploiting modern cloning methods. The new vectors build on the existing pMTL8000 vectors that have been a staple of Clostridium research for more than a decade. The introduction of two pairs of type IIS restriction sites flanking an insulated multiple cloning site in both a cloning vector and a CRISPR-Cas9 gene editing vector enables plasmid construction in a “one-pot” reaction, avoiding the more laborious steps of conventional cloning. A synthetic lacZα expression cassette introduced between the cloning sites enables visual detection of background colonies. In addition, distinct selection markers on each vector permit selection of the desired clones according to antibiotic resistance. An example of strain development using the new vectors is demonstrated. |
| first_indexed | 2024-03-12T15:30:08Z |
| format | Article |
| id | doaj.art-05e2037815a9408fb021a3074f056575 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-03-12T15:30:08Z |
| publishDate | 2023-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-05e2037815a9408fb021a3074f0565752023-08-10T04:34:49ZengElsevieriScience2589-00422023-08-01268107484Streamlined assembly of cloning and genome editing vectors for genus ClostridiumTom S. Bailey0Philip Hittmeyer1Yanchao Zhang2Aleksandra M. Kubiak3GROW - School for Oncology and Reproduction, Faculty of Health and Medical Life Sciences, Maastricht University, Maastricht, Limburg, the Netherlands; Corresponding authorGROW - School for Oncology and Reproduction, Faculty of Health and Medical Life Sciences, Maastricht University, Maastricht, Limburg, the NetherlandsGROW - School for Oncology and Reproduction, Faculty of Health and Medical Life Sciences, Maastricht University, Maastricht, Limburg, the NetherlandsExomnis Biotech BV, Oxfordlaan, Maastricht, Limburg, the NetherlandsSummary: Reported herein is a new set of vectors designed to streamline molecular cloning and genome editing by exploiting modern cloning methods. The new vectors build on the existing pMTL8000 vectors that have been a staple of Clostridium research for more than a decade. The introduction of two pairs of type IIS restriction sites flanking an insulated multiple cloning site in both a cloning vector and a CRISPR-Cas9 gene editing vector enables plasmid construction in a “one-pot” reaction, avoiding the more laborious steps of conventional cloning. A synthetic lacZα expression cassette introduced between the cloning sites enables visual detection of background colonies. In addition, distinct selection markers on each vector permit selection of the desired clones according to antibiotic resistance. An example of strain development using the new vectors is demonstrated.http://www.sciencedirect.com/science/article/pii/S2589004223015614Molecular GeneticsBiotechnologyMicrobial biotechnology |
| spellingShingle | Tom S. Bailey Philip Hittmeyer Yanchao Zhang Aleksandra M. Kubiak Streamlined assembly of cloning and genome editing vectors for genus Clostridium iScience Molecular Genetics Biotechnology Microbial biotechnology |
| title | Streamlined assembly of cloning and genome editing vectors for genus Clostridium |
| title_full | Streamlined assembly of cloning and genome editing vectors for genus Clostridium |
| title_fullStr | Streamlined assembly of cloning and genome editing vectors for genus Clostridium |
| title_full_unstemmed | Streamlined assembly of cloning and genome editing vectors for genus Clostridium |
| title_short | Streamlined assembly of cloning and genome editing vectors for genus Clostridium |
| title_sort | streamlined assembly of cloning and genome editing vectors for genus clostridium |
| topic | Molecular Genetics Biotechnology Microbial biotechnology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004223015614 |
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