Skip to main content
. 2004 Oct;186(19):6465–6476. doi: 10.1128/JB.186.19.6465-6476.2004

FIG. 3.

FIG. 3.

(A) SDS-PAGE profile of purified recombinant 17-kDa protein. The proteins were resolved on an SDS—12% PAGE gel and were visualized by staining with Coomassie brilliant blue or blotted with antiserum. Lane 1, purified recombinant 17-kDa protein without histidine tag after DEAE ion-exchange purification; lane 2, Ni-NTA affinity column-purified 17-kDa protein; lane 3, Ni-NTA affinity column-purified triple-mutant protein (Val-Phe-Ile → Ala-Ala-Ala). Numbers on the left are molecular weight markers (in thousands). (B) Western blot of recombinant 17-kDa pilin protein of X. nematophila. Lanes 1, 2, and 3 were developed with monoclonal antibodies against the six-His tag. Lanes 4, 5, and 6 were developed with polyclonal antibodies against native 17-kDa protein of X. nematophila. Lane 1, cell lysate of uninduced culture of strain PK5; lane 2, cell lysate of induced culture of strain PK5; lanes 3 and 5, Ni-NTA-purified 17-kDa protein; lane 4, DEAE column-purified 17-kDa protein (without tag); lane 6, Ni-NTA-purified triple-mutant protein (Val-Phe-Ile → Ala-Ala-Ala). Numbers on the left are prestained markers.