FIG. 5.
B-band LPS production can be restored by a wzyβ gene, but not by a wzyα gene, in a wzyβ knockout mutant. (A) Silver-stained SDS-PAGE gel of LPS from serotype O5 (wild type), serotype O2 (wild type), serotype O16 (wild type), an O16 wzyβ knockout mutant, an O16 wzyβ mutant transformed with pUCP27 (control), an O16 wzyβ mutant complemented with O2 wzyβ, and an O16 wzyβ mutant complemented with O2 wzyα. Note that the banding modality of the O16 wzyβ mutant complemented with O2 wzyβ is very similar to that of wild-type O16, indicating that S-LPS was being restored. O2 wzyα could not restore S-LPS in the O16 wzyβ mutant strain, except for one HMW band, which is indicated by an arrow. (B) Western blots of the samples in panel A reacted with O16-specific MAb. O16-specific MAb reacts with LPS from the O16 wzyβ mutant complemented with O2 wzyβ, but not with LPS from the O16 wzyβ mutant complemented with O2 wzyα, indicating that wzyα cannot restore B-band production in the O16 wzyβ knockout mutant. (C) Western blots of the samples in panel A reacted with O5-specific MAb. O5-specific MAb can react only with α-linked O antigen, which in this blot was found exclusively in the LPS from O5.