secretion of lysosomal proteins?

Robert Kuhelj robert.kuhelj at ijs.si
Sat Feb 4 13:58:33 EST 1995


Hi,

I'd like to express a human lysosomal protein in a mammalian and/or
baculoviral system.

If the protein's native signal sequence is included when constructing
the expression vector, the protein will probably be targeted to 
lysosomes. To isolate it, I'd have to break the cells, which I would
like to avoid!

Is there any way to make the cells secrete lysosomal proteins into the medium?
In this way, cells wouldn't have to be lysed, the recombinant protein
production would be continuous. I didn't find any commercial expression
vector with an appropriate signal sequence to target the protein out of the
cell (heterologous  cDNA could then  be inserted directly downstream,
without disrupting the ORF, of course).

Are there any additives whose presence in the culture medium would change 
the targeting in a way that I have described?  
_______________________________________________________________________________
Robert Kuhelj                 Jozef Stefan Institute
                              Department of Biochemistry and Molecular Biology
                              Ljubljana
robert.kuhelj at ijs.si          Slovenia
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