Hi there,
Somehow I've missed the original email :) Sorry!
There are options for expressing really toxic genes, some of which have
already been mentioned and others perhaps not:
1. tight regulation of expression (promoter, repressor, other regulatory
elements, or a combination thereof). Beyond
Hi Andy,
just to follow up on Christian suggestion, which is exactly the way to go.
In case you are using an already pET based vector, simply try BL21-AI
(https://www.thermofisher.com/order/catalog/product/C607003), which has the T7
RNA polymerase under arabinose promoter should do the trick.
Hi Andy,
have you tried another promotor? Arabinose is much tighter, just to be sure
that it is really not leaking.
Cheers
Christian
On Mon, Apr 4, 2022 at 10:20 AM Andrew Lovering
wrote:
> Dear Board,
>
>
>
> Perhaps off-topic, but in the wider scope it’s relevant to many on here.
>
>
>
> We
Dear Board,
Perhaps off-topic, but in the wider scope it's relevant to many on here.
We have a gene that we are able to clone, and propagate in DH5a etc
non-expression cells (hence nucleotide sequence is non-toxic)
But, when we attempt to transfer to an expression strain we get no colonies
We
