FIG. 3.

Cloning, expression, and purification of rMYPE9110. (A) Schematic representation of seven TGA codons corrected to TGG codons. Six overlapping fragments were amplified with the mutagenic primers listed in Table 1 and annealed in order to change UGA-encoding tryptophans to UGG-encoding tryptophans and generate a full-length MYPE9110 gene product to express in E. coli cells. (B) Expression and purification of rMYPE9110. rMYPE9110 was expressed and purified in the E. coli BL21(DE3) lpxM strain, resolved by 10% Laemmli SDS-polyacrylamide gel electrophoresis, and stained with Coomassie brilliant blue R250. Lane 1, uninduced total cell lysate; lane 2, induced total cell lysate; lane 3, purified rMYPE9110.