Quick navigation:        Home   |    Site Map   ||    References   |    Biography   ||    Copyright   |    Other copyright   |    Contact us   |   
Protein structure
 

Re: [ccp4bb] off topic - C-ter cleavable his tag
 

Basic tutorials: 		 
 

CCP4bb navigation

CCP4bb <-- 2008 <-- April 2008 <-- 04 April 2008
Previous message:
Subject: off topic - C-ter cleavable his tag
From: Vijay Kumar vijay85 {- at -} GMAIL {- dot -} COM
Date: 2008-04-04 		Next message:
Subject: Re: Off topic: McDonalds Happy Meal and Crystallography
From: Ezra Peisach epeisach {- at -} BU {- dot -} EDU
Date: 2008-04-04


Subject: Re: off topic - C-ter cleavable his tag
From: Chun Luo chun {- at -} ACCELAGEN {- dot -} COM
Date: 2008-04-04
Hi Vijay,



You may consider having a protease site in your 3' PCR primer, since you
probably have to remove the stop codon at the same time. It's probably
easier to do than finding a vector with C-terminal cleavable tag and C-His
will be in-frame without any extra sequence. Many pET vectors with N-His
also have C-His. You may also consider HRV3C or TEV. Not because Accelagen
sells those proteases, but they are more specific than proteases like
thrombin and easier to remove.



Cheers,



Chun



Chun Luo, Ph.D.
The Protein Expert
Accelagen, Inc.
11585 Sorrento Valley Road, Suite 107
San Diego, CA 92121
TeL: 858-350-8085 ext 111
Fax: 858-350-8001
chun@accelagen.com
www.accelagen.com





_____

From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Vijay
Kumar
Sent: Friday, April 04, 2008 8:40 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] off topic - C-ter cleavable his tag



Hi,

I am looking for a vector with a cleavable C-terminal his tag for
prokaryotic protein expression. I found pET-52b but it has also got a
N-terminal cleavable strep-tag which I cannot avoid.

Thanks

Vijay
ProteinCrystallography.org: Copyright 2006-2008 by Quid United Ltd