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. 2007 Nov 5;76(1):417–425. doi: 10.1128/IAI.00986-07

FIG. 4.

FIG. 4.

Expression and purification of PCRan1p and PCMei2p. A. S. cerevisiae strain INVSC− with PCRan1 was cultured in 1.5 liters of dropout base medium lacking uracil and containing 2% galactose to induce expression of the transgene for 24 h. The protein was purified by His-affinity using a nickel-charged column and collected into five separate fractions. These fractions were separated on a 12% SDS-PAGE and Western blotted using the specific PCRan1p antibody. B. Similarly, S. cerevisiae strain INVSC− with PCMei2 was cultured, and expression of the recombinant protein was induced as described above. Culture supernatants were purified as described above and collected into five separate fractions. These fractions were separated on a 12% SDS-PAGE and Western blotted using an anti-V5 antibody.