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Site-specific mutagenesis. The p35 intron was eliminated by site-directed mutagenesis. Briefly, we inserted a BamHI restriction site (underlined) in the three codons (encoding Arg, Gly, Ser) to the right of the intron. Primers were designed to PCR amplify two fragments of p35 that were joined at the BamHI site, such that p35 was reconstituted but intron free.
