BL21(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.
BL21(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.
BL21(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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| Strain | Escherischia coli |
| Delivery condition | The strain is provided on a lysogeny broth (LB) agar plate containing 20 µg/mL chloramphenicol. |
| Storage temperature | 4 °C |
Publication
gger, E., Tauer, C., Cserjan-Puschmann, M., Grabherr, R. & Striedner, G. Fast and antibiotic free genome integration into Escherichia coli chromosome. Sci. Rep. 10, (2020).
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