I agree. I think it depends on the lab and vectors and personal preference.
But David, have you used them sequentially? I have once tried to make a His-GST-ENLYFQ-3C construct, and I found that it was self cleaving during expression. However I have never tried to replicate the results in vitro. -- Kelvin Lau Protein production and structure core facility - PTPSP EPFL SV PTECH PTPSP AI 2146 (Bâtiment AI) Station 19 CH-1015 Lausanne Switzerland Email: [email protected]<mailto:[email protected]> Phone: +41 21 69 34494 On 7 Dec 2022, at 21:38, David Briggs <[email protected]<mailto:[email protected]>> wrote: Hi Gloria, Both can be made very easily in E.coli. Both are active at 4°C, but especially 3C, I think. I have plasmids for both somewhere in the freezer (you might find someone closer to you who can send HRV3C, but if you cannot, let me know off list). I don't see any particular benefit of one over the other, but having both in your freezer means you can cleave off tags sequentially as needed by your purification strategies. HTH, Dave Dr David C. Briggs CSci MRSB Principal Laboratory Research Scientist Signalling and Structural Biology Lab The Francis Crick Institute London, UK == about.me/david_briggs<http://about.me/david_briggs> ________________________________ From: CCP4 bulletin board <[email protected]<mailto:[email protected]>> on behalf of Gloria Borgstahl <[email protected]<mailto:[email protected]>> Sent: Wednesday, 7 December 2022, 20:26 To: [email protected]<mailto:[email protected]> <[email protected]<mailto:[email protected]>> Subject: [ccp4bb] TEV vs HRV3C External Sender: Use caution. Hello my fellow structural biologists, I am contemplating why some choose the HRV3C protease site over TEV for their fusion proteins. Does anyone know? Can HRV3C be made easily in homelab? Does anyone have a plasmid? Thank you, G ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1<https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=05%7C01%7C%7Cb865bd537cfa41af3b0f08dad8914aeb%7C4eed7807ebad415aa7a99170947f4eae%7C0%7C1%7C638060415788656699%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000%7C%7C%7C&sdata=M1H4lHtUxJ1WZR0%2B73eYsZCc8%2FBDO8pWcV0yjFgomHk%3D&reserved=0> The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT ________________________________ 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/
