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Re: [ccp4bb] Tag, you're in! Flag, V5, etc |
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CCP4bb navigationCCP4bb <-- 2008 <-- February 2008 <-- 07 February 2008Subject: Re: Tag, you're in! Flag, V5, etc From: James Whisstock James {- dot -} Whisstock {- at -} MED {- dot -} MONASH {- dot -} EDU {- dot -} AU Date: 2008-02-07 We usually add all N-terminal tags with a TeV cleavable linker. C-terminal tags always seem a pain to remove cleanly, because most highly specific recognition sequences (such as TeV) take advantage of P rather than P' specificity so you are usually left with a five (or so) residue tail. Actually has anyone any neat tricks for C-terminal tag removal? J "Y. -F. Li" > Hello, > > I'd appreciate it if anyone could provide information (experiences or > publications) on the following: > > 1. Put a tag such as FLAG, V5, etc etc, at the N- and/or C-termini, in > order > for specific detection, but not interfering with protein > folding/structure; > 2. Is a linker between the tag and target protein needed? What linkers > (length, specific sequences) would you suggest? > 3. What are the choices of such tags and why would you recommend them? > > Thank you for your time and sharing in advance. > > Yongfu Li -- Professor James Whisstock NHMRC Principal Research Fellow / Monash University Senior Logan fellow Department of Biochemistry and Molecular Biology Monash University, Clayton Campus, PO Box 13d, VIC, 3800, Australia +613 9905 3747 (Phone) +613 9905 4699 (Fax) +61 418 170 585 (Mobile) CCP4bb navigationCCP4bb <-- 2008 <-- February 2008 <-- 07 February 2008 |
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