Glucocorticoid-inducible vector troubles
a.plume at botany.uq.edu.au
Wed Sep 26 00:37:12 EST 2001
I have produced homozygous, single-copy lines containing either the
empty pTA7001 glucocorticoid-inducible vector (Aoyama and Chua, 1997) or
with my genes of interest inserted in the sense orientation at the
polylinker. I have selected one line of each and shown that the GVG
levels are equivalent at the RNA level and have been successfully able
to induce overexpression at the RNA level of my genes of interest, but
cannot see if there is an associated phenotype because of my problem.
My problem is this: After about 1 week on the inducer (10uM
Dexamethasone, DEX) my plants go yellow and die. This occurs in the
empty vector line as well as in my experimental lines. I have two ideas
about the cause:
1) Is it an impurity in the Dex?
2) Is the GVG expression itself too high, causing cell death?
Any ideas greatly appreciated.
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