Hi,
I am interested in using a regulatable tRNA suppressor gene, to
create conditional mutants in my favorite gene cerevisiae. I have
been unable to generate ts mutants of my gene, and unable to confer
tight transcriptional regulation on it using the various regulatable
promoters that are available (they are too leaky). So I thought I
might be able to engineer an amber (f'rinstance) mutation at the
beginning of my ORF and then conditionally turn it on by upregulating
SUP3 (f'rinstance). So does anybody have or know where I can get
such a reagent? Can you generate a functional tRNA by putting it
under control of an RNA Pol II promoter? I'm considering putting
SUP4 or SUP11 under GAL control and seeing what happens. Please help!
bd033054 at condor.mbcr.bcm.tmc.edu
