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. 1999 Feb;67(2):675–680. doi: 10.1128/iai.67.2.675-680.1999

FIG. 2.

FIG. 2

Expression and purification of recombinant antigens of pypag-1. (A) Diagram showing the sequences of pypag-1 PCR amplified by using primers A through D and ligated into the pET-15b bacterial expression vector. Recombinant pAg-1N contains amino acids 1 to 337 of pypAg-1, including a potential membrane-spanning sequence (black box). Recombinant pAg-1C contains amino acids 339 to 549 of pypAg-1, including the repeat domain (hatched box). N and X represent NdeI and XhoI restriction sites introduced to facilitate subcloning. (B) Coomassie blue-stained SDS-polyacrylamide gel containing E. coli lysates of uninduced (lane 1) and induced (lane 2) cells expressing pAg-1N, purified pAg-1N (2.5 μg, lane 3), E. coli lysates of uninduced (lane 4) and induced (lane 5) cells expressing pAg-1C, and purified pAg-1C (2.5 μg, lane 6). Molecular mass markers are indicated.