Protein expression and purification Bacteria - Escherichia coli GmrSD
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Discussion
5 years ago
5 years ago by
Dobrogost Maciejewski
What temperature should I use for expressing thermophillic genes in E. coli?
I am currently working with E.coli and I am trying to express thermophilic genes. What temperature should I be using for incubation after IPTG induction? I have read some papers that suggested to incubate the cells at 42 degrees for a short amount of time. Is this going to affect the protein?
Discussion
5 years ago
5 years ago by
Lalli Rislakki
Can IPTG in alkaline conditions become toxic to the bacteria?
Hi everyone! Currently I am attempting to measure the beta -galactosidase activity by inducing the lac operon in E.coli cells. I am trying to grow my cells in alkaline conditions with a pH > 8.5 but when I induce the cells with IPTG they are not growing. The IPTG concentration I have tried using is 0.1 and 1mM. Is there a chance that IPTG in these concentrations in alkaline conditions becomes toxic to my bacteria? Should I lower the concentration or do you think that the problem lies elsewhere?
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