Promega is selling those Flexi-System Vectors. However, they come with Barnase as a selection marker and hence cannot simply be transformed into your usual library-competent E. coli strain. Since I do not want to buy purified plasmid every time I run out of it I am looking for an option to propagate those plasmids myself. Does anyone have a barstar-expressing E.coli strain that could be transformed with barnase expressing plasmids? They should be compatible with the ColE1/pMB1/pBR322/pUC-type ori if the barstar-cassette is on a plasmid.
You can get information from this reference-
Barnase and barstar. Expression of its cloned inhibitor permits expression of a cloned ribonuclease. Hartley RW. J Mol Biol 1988 Aug 20;202(4):913-5. 10.1016/0022-2836(88)90568-2 PubMed 3050134
Hello Christoph,
this group from New Zealand have worked with Barstar-expressing E.coli strain. Maybe they can provide you the plasmids you need for E. coli transformation.
Article Exploring the parameters of post segregation killing using h...
Hope it helps.
In case this is still of interest:
Barnase/Barstar can be obtained in a single plasmid here:
https://www.addgene.org/8606/
This plasmid originates from the above mentioned publication
Barnase and barstar. Expression of its cloned inhibitor permits expression of a cloned ribonuclease. Hartley RW. J Mol Biol 1988 Aug 20;202(4):913-5. 10.1016/0022-2836(88)90568-2 PubMed 3050134
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