Re: [ccp4bb] Off-topic: Duet vectors with C-terminal GST tag
How about just co-transfecting with two different plasmids which have different selection markers? I've done it a lot, and it seems to work fine... JPK On Fri, Aug 24, 2012 at 10:39 PM, Lye, Ming ming_...@hms.harvard.eduwrote: Dear CCP4bb, We would like to co-express proteins under Se-Met conditions for de-novo phasing of a complex. One of the proteins expresses much better with a GST-tag compared with a his-tag. As its binding site is at its N-terminus, we are hoping to co-express it as a C-terminal GST tag recombinant. So far however, we haven't found any commercially available duet vectors with such a tag. We would truly appreciate if anyone knows of the availability of such vectors, or has any suggestions on similar vectors. Thanks so much! Ming -- *** Jacob Pearson Keller Northwestern University Medical Scientist Training Program email: j-kell...@northwestern.edu ***
Re: [ccp4bb] Off-topic: Duet vectors with C-terminal GST tag
Hi Jacob, Thanks for your message! I was thinking about that too, but I wasn't sure if it was something that just sounded good in theory.. I guess the two different vectors should have different antibiotic resistance to select for the 'co-transformation.' Thanks so much! Ming From: Jacob Keller [j-kell...@fsm.northwestern.edu] Sent: Monday, August 27, 2012 9:31 AM To: Lye, Ming Cc: CCP4BB@jiscmail.ac.uk Subject: Re: [ccp4bb] Off-topic: Duet vectors with C-terminal GST tag How about just co-transfecting with two different plasmids which have different selection markers? I've done it a lot, and it seems to work fine... JPK On Fri, Aug 24, 2012 at 10:39 PM, Lye, Ming ming_...@hms.harvard.edumailto:ming_...@hms.harvard.edu wrote: Dear CCP4bb, We would like to co-express proteins under Se-Met conditions for de-novo phasing of a complex. One of the proteins expresses much better with a GST-tag compared with a his-tag. As its binding site is at its N-terminus, we are hoping to co-express it as a C-terminal GST tag recombinant. So far however, we haven't found any commercially available duet vectors with such a tag. We would truly appreciate if anyone knows of the availability of such vectors, or has any suggestions on similar vectors. Thanks so much! Ming -- *** Jacob Pearson Keller Northwestern University Medical Scientist Training Program email: j-kell...@northwestern.edumailto:j-kell...@northwestern.edu ***
[ccp4bb] Off-topic: Duet vectors with C-terminal GST tag
Dear CCP4bb, We would like to co-express proteins under Se-Met conditions for de-novo phasing of a complex. One of the proteins expresses much better with a GST-tag compared with a his-tag. As its binding site is at its N-terminus, we are hoping to co-express it as a C-terminal GST tag recombinant. So far however, we haven't found any commercially available duet vectors with such a tag. We would truly appreciate if anyone knows of the availability of such vectors, or has any suggestions on similar vectors. Thanks so much! Ming
