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gain of function mutagenesis, how to do?

Yoram Gerchman yoramg2 at leonardo.ls.huji.ac.il
Sun Jan 30 05:43:48 EST 2000


Dear Ricky
I would go for random mutagenesis. If you have 30% difference you can
try and mutate these region using the Mega prime method. Make a
mutagenic PCR on the region of interest and use the PCR product as a
primer for another, non-mutagenic PCR reaction. You will still need some
cloning...
Are you might want to try chemical mutagenesis with Hydroxyl Amid (It
only change C->T on both strands).
Again it might be easier to mutate the working gene and look for loss of
activity. A "bad" mutation is more likely
Regards Yoram
.........................................................................
"Support bacteria - they're the only culture some people have"
                                     (anonymous)
Yoram Gerchman
Division of Microbial & Molecular Ecology
The Institute of Life Sciences
Hebrew University of Jerusalem
Givat Ram, Jerusalem 91904,
Israel
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