| Quick navigation: | Home | Site Map || References | Biography || Copyright | Other copyright | Contact us | | |
|
|
Re: [ccp4bb] Co-expression plasmids |
|
CCP4bb navigationCCP4bb <-- 2008 <-- April 2008 <-- 02 April 2008Subject: Re: Co-expression plasmids From: P Hubbard dienoylcoa {- at -} HOTMAIL {- dot -} COM Date: 2008-04-02 Just to explain to people who've never had to do co-expression - if I remember correctly, conventional wisdom states that you can't have two different vectors with the same origin of replication in the same host (maybe someone might know more details as to why). However, as pointed out, it can be made to work. Alas, not in my case! CC: CCP4BB@JISCMAIL.AC.UK From: a.perrakis@nki.nl Subject: Re: [ccp4bb] Co-expression plasmids Date: Wed, 2 Apr 2008 19:59:09 +0200 To: dienoylcoa@HOTMAIL.COM We have had good experience with the awfully simple minded approach of using two pET vectors with different antibiotic resistance.Its the easiest thing to do, and it often works ... Apologies for the shameless plugin, since there are many good papers on the subject, but you can read some hints and case studies at: http://scripts.iucr.org/cgi-bin/paper?S0907444906031003 (its Open Access) A. On 2 Apr 2008, at 19:39, P Hubbard wrote:Hi, If you are expressing just two proteins, you could try a single pET vector with a pCDF vector. The only reason I'm suggesting this is that I had trouble with pET-Duet, but doing each one separately worked first time (plasmid size issue?). I used pET24-a and pCDF-1b - so I had one construct untagged, and the other with a cleavable His-tag. Cheers AGS Date: Wed, 2 Apr 2008 15:55:27 +0200 From: VANRAAIJ@USC.ES Subject: [ccp4bb] Co-expression plasmids To: CCP4BB@JISCMAIL.AC.UK Dear All,Anyone have experience with the NovaGen Duet co-expression vectors? Or can recommend others? http://www.emdbiosciences.com/html/NVG/Duet_Spot.html Greetings,MarkMark J. van RaaijDpto de Bioquímica, Facultad de FarmaciaUniversidad de Santiago15782 Santiago de CompostelaSpainhttp://web.usc.es/~vanraaij/ Get in touch in an instant. Get Windows Live Messenger now. _________________________________________________________________ Get in touch in an instant. Get Windows Live Messenger now. http://www.windowslive.com/messenger/overview.html?ocid=TXT_TAGLM_WL_Refresh_getintouch_042008 CCP4bb navigationCCP4bb <-- 2008 <-- April 2008 <-- 02 April 2008 |
| ProteinCrystallography.org: Copyright 2006-2008 by Quid United Ltd |