Vectors for mammalian expression and secretion of recombinant
nobody at nospam.not
Thu Dec 22 20:25:17 EST 2005
"troels.wind at gmail.com" <troels.wind at gmail.com> wrote in
news:1135260572.478683.126910 at f14g2000cwb.googlegroups.com:
> Dear all,
> Does anyone know of a vector for expressen in e.g. HEK293 similar to
> pcDNA but with a versatile secretion signal upstream of the multiple
> cloning site? We wish to express and secrete recombinant proteins
> without an "inherent" secretion signal.
> Thanx and merry xmas
>From our paper in JCI <http://www.jci.org/cgi/reprint/101/9/1851>
R. B. Gayle, III, C. R. Maliszewski, S. D. Gimpel, M. A. Schoenborn, R.
G. Caspary, C. Richards, K. Brasel, V Price, J. H. F. Drosopoulos, N.
Islam, T. N. Alyonycheva, M. J. Broekman, and A. J. Marcus. Inhibition of
platelet function by recombinant soluble ecto-ADPase/CD39. J.Clin.Invest.
101 (9):1851-1859, 1998.
> To allow proper transport of soluble CD39 to the medium, an appropriate
> leader sequence providing for secretion was required at the amino
> terminus of the protein. Thus, the amino terminal transmembrane region
> was replaced by a leader derived from human proinsulin/IL-2 sequences
> (ref 15) (IL-2L) and a sequence coding for a small peptide tag, Flag.
J. H. F. Drosopoulos, M. J. Broekman, N. Islam, C. R. Maliszewski, R. B.
Gayle, III, and A. J. Marcus. Site-directed mutagenesis of human
endothelial cell ecto-ADPase/soluble CD39. Requirement of glutamate174
and serine218 for enzyme activity and inhibition of platelet recruitment.
Biochemistry 39 (23):6936-6943, 2000.
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