Figure 5

PCR amplification for multiple-site mutagenesis. A) Agarose gel electrophoresis of the PCR reactions indicating the amplification efficiency. The names of the mutants are shown on the top of each lane. B) Transformation and mutation efficiency for 3026L/M-51L/M, both Leu26 and Leu51 in CAG38830 substituted by methionines, 3327L/M-56L/M, both Leu27 and Leu56 in CAG38833 substituted by methionines, VraRDNH/ICH, a cloned vraR gene with its N-terminal His tag removed and C-terminal His tag inserted and VraRDN3/DC5, a cloned vraR gene with three residues from the N-terminus and five from the C-terminus deleted. Arrows indicate the partial PCR amplification products.