Fig. 1

Construction of tagged mutant p53 expression plasmids. Plasmids were constructed in a three step process. (A) 200 bp tags containing six variable positions were synthesized using overlapping oligonucleotide PCR; tags were cloned into the expression plasmid using homologous recombination mediated gap repair. (B) Mutant p53 genes were constructed using mutagenic crossover PCR; both degenerate (–NNN–) and mutant specific primers were used. (C) Final plasmid assembly was done by ligating SpeI/HindIII digested tagged vector and mutant p53 genes from (A) and (B)