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 <a.lover...@bham.ac.uk> wrote: > 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 have tried pLemo, Glucose addition, 30 degrees, C41/C43 and still no joy > > > > We’d welcome any suggestions here – it’s a fun protein > > > > Thanks > > Andy > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
