Sebastien.Paris at UNIV-ROUEN.FR (Sebastien Paris) wrote:
>I'm using the bicistronic pIRES-hyg vector to obtain stable clones for a
>protein of interest. Is it possible to obtain only resistant clones to hyg
>which do not express the protein of interest. If it's possible, has anybody
>a suggestion to obtain a clone which express this protein ?
It is possible if:
a. your insert contains cryptic splice sites so that the coding
sequence - or part of it - is lost;
b. if there is a rearrangement within the vector (I had approx.
10-15% rearrangement with no expression using retroviral vectors),
for plasmid vectors it should be a rare event.
The only way you can be ABSOLUTELY sure is to test for the presence of
the product :-).
Victor Levenson MD/PhD
Res Asst Prof
UIC, Dept of Genetics M/C 669
900 S Ashland Ave The one who wants finds HOW
Chicago IL 60607 The one who doesn't finds WHY
levenson at uic.edu