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 <mailto:[EMAIL PROTECTED]> [EMAIL PROTECTED] www.accelagen.com <http://www.accelagen.com/> _____ From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Vijay Kumar Sent: Friday, April 04, 2008 8:40 AM To: [email protected] 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
