HMS174(DE3)::I-SceIRS (pAIO)
Escherichia coli host strain carrying the I-SceI restriction site on genome and the pAIO plasmid for antibiotic free genome integration on attTN7 site.
HMS174(DE3)::I-SceIRS (pAIO) offers a I-SceI homing endonuclease restriction site on genome. The 18 nucleotides long sequence was inserted at the attTN7 site and allows for selection during pAIO-based genome integration.
HMS174(DE3)::I-SceIRS (pAIO) additionally carries the pAIO plasmid which is equipped with all elements for “All-In-One” genome integration. The λ-red enzymes Exo, Gam and Bet under control of a heat inducible promotor were selected to ensure the integration of a linear target gene cassette into the E. coli genome. The gene encoding the homing endonuclease I-SceI was inserted into the vector under control of the araB promoter for selection purpose and plasmid curing. To provide accelerated plasmid curing, we engineered an additional I-SceI site into the plasmid backbone, with the idea that the plasmid also gets cleaved upon the expression of I-SceI (doi: 10.1038/s41598-020-73348-x).
Material for Research and Development purposes only.
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| Storage temperature | 4 °C |
| Delivery condition | The strain is provided on a lysogeny broth (LB) agar plate containing 20 µg/mL chloramphenicol. |
| Strain | Escherichia coli |
Publication
Fink M, Vazulka S, Egger E, Jarmer J, Grabherr R, Cserjan-Puschmann M, Striedner G. Microbioreactor Cultivations of Fab-Producing Escherichia coli Reveal Genome-Integrated Systems as Suitable for Prospective Studies on Direct Fab Expression Effects. Biotechnol J. 2019 Nov;14(11):e1800637. doi: 10.1002/biot.201800637. Epub 2019 Jul 26. PMID: 31231932.
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