Mutagenesis allowed to obtain PAP1 fragment without inconvenient sequences that could interact with restriction enzymes and interfere with the later insertion into plasmids. (Fig. 3A)
Following this mutagenesis, we obtain the small fragments of PAP1 that need to be reassembled. The reassembly is done again by PCR. The control is done on a gel where we check if we obtain the same size as for the original PAP1. Here we can clearly see that reassembly of new mutated PAP1 fragments gives the same final size as for the original one(Fig. 2B).
Plasmid insertion of PAP1 to 11