Hi, I am working with the Tet-repressor in tobacco to express a gene that
might be lethal to plants. I got a lot of transformants with my constructs
but was not able to detect a protein in Western Blots of induced leaves.
Because the induction of a control with GUS was not very good in leaf I
started supension cultures. I got good induction of the GUS construct in
the suspensions and was able to detect the §-Glucuronidase in a Western
Blot after induction. Nevertheless I never saw any sighn of the Protein I
am interested in.
Due to John Shanklin I am now suspisious that it might not be my Protein
that is the problem but the system. Therefore I would like to know if
anybody else got the system to work in any plant with a Protein that is not
GUS?? Did you check for RNA?