In my experience, 3C can partially cut TEV sites as well. If using sequentially, it is best to plan to use the TEV first.
> On Dec 7, 2022, at 2:42 PM, Lau Kelvin > <[email protected]> wrote: > > 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 >> <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 > <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/
