Help Needed for Oocyte RNA Expression Study

RiverSloth riversloth at aol.com
Mon Nov 21 16:00:11 EST 1994


I am about to send some in vitro transcribed RNA to collaborators at
another location where they will microinject the RNA into Xenopus oocytes.
We hope to get the RNA translation product incorporated into the oocyte
cell for receptor binding assays-the RNA codes for a neuropeptide hormone
receptor.

Since oocyte expression studies are a new area of study for me, I was
hoping to get some advice on a couple of questions I have:

1.  Will the length of the 5' untranslated region of the RNA affect the
translatibility of injected RNA in the oocyte system?

2.  Will spurious AUG codons occuring 5' of the "correct" AUG codon for
the receptor protein open reading frame prevent translation of the correct
open reading frame, particularly if the spurious AUGs have termination
codons? Or will the ribosomes resume scanning the RNA for the next AUG,
eventually reaching the receptor open reading frame?

I appreciate any help you can give me. I am trying to anticipate problems
before they arise. I expect any problems with the assay will be laid at
the feet of the RNA supplier (me!) so I hope to be ready with some
answers.

Thanks,

Felix D. Guerrero, PhD
USDA-ARS
Kerrville, TX



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