omission mutagenesis

[Bruce D. Erickson]

In article <3vmdgd$d9d at mark.ucdavis.edu> szeliasf at chip.ucdavis.edu (Elias Fernandez) writes:

>would anyone have carried out a mutagenesis where a single residue was 
>ommited in the coding gene? 
>would it be possible to use a primer with a codon missing and generate 
>the mutant by pcr amplification so that the protein has a residue missing?

>thanks
>elias

I haven't tried it with PCR, but it worked just fine using an 
oligonucleotide-mediated mutagenesis system (in my case the Transformer system 
form Clontech).  I used a 20-mer oligo with ten nucleotides on either side of 
the three nucleotide gap.  I don't recall what the efficiency was, but I got 
the clone in the first set of mutants that I screened.

Good Luck

Bruce

(No affiliation with Clontech)

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