Figure 2.

Site-directed mutagenesis. (A) Strategy to introduce targeted mutations into target genes using two-rounds of PCR. Primers used to introduce cut14 hinge mutations are presented in Table S1 and primers used to introduce cut3 hinge mutations are presented in Table S2. (B) Site-directed mutagenesis and isolation of ts/cs mutants. See ‘Materials and Methods’ for detailed description of plasmid construction and how to isolate ts/cs mutants.