c-terminal tag for transmembrane protein

Francisco G da Nobrega ib - bio 7588 fgdnobre at usp.br
Fri Oct 25 09:00:39 EST 1996


Dear Robin:

I learned with Dr. Alexander Tzagoloff to tag mitochondrial membrane 
proteins with a biotin tag. The suggestion is from the work of Cronan 
J Biol Chem 265:10327-33 (1990)
and Alex prepared a number of plasmids to make it easier to construct 
derivatives. We usually make a translational fusion of the 
bacterial transcarboxylase biotination peptide to the C terminal end of 
the protein. Most constructions retain the biological activity and the 
addition of the biotin group is done in vivo in the yeast cell.
Dr. Tzagoloff is at the Biological Sciences Dept., Columbia University, 
New York City

F.G. Nobrega
Universidade de Sao Paulo
Departamento de Microbiologia - ICB II / USP
Av. Prof. Lineu Prestes, 1374
05508-900   Sao Paulo  -  SP  - Brasil
Voice: (011) 818-7588    Fax: 55-11-818-7354    E-mail:  fgdnobre at usp.br

On 24 Oct 1996 rtemple at netmail.hscbklyn.edu wrote:

> Hello all -
> 
> I've got a 12-pass channel protein that I'd like to tag & visualize. I don't
> want to mess w/ any signaling on the amino end, and internal tags could mess up
> function. After checking the catalogs, I don't see a vector around that will
> carboxy-tag commercially and I don't want to 'hand clone' repetitive HA's on the
> end unless I have to do so. I do not want to raise Ab's to an integral protein.
> Does anyone know of a source of such a vector? Any help would be greatly
> appreciated. 
> 
> Thanks in advance -
> Robyn Temple
> SUNY Health Science Center at Brooklyn
> 
> 
> 



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