1887

Abstract

The / gene, encoding the O-antigen polymerase, of serovar Typhimurium has been previously cloned and sequenced. In the present work, the transcriptional startpoint was initially identified by primer extension. Next, promoter strength in K-12 was measured, and was found to be greater than that of the induced promoter. To define the Wzy translational startpoint, DNA including the promoter and the putative first five residues of the Wzy protein was fused to the N-terminus of glutathione-S-transferase, and the fusion protein purified by affinity chromatography. N-terminal amino acid sequencing yielded the Wzy translational startpoint. Next, the Wzy protein was C-terminally tagged with the FLAG peptide, and immunoblotting of an strain expressing a low-copy gene (five copies per cell) localized the intact Wzy protein in the cytoplasmic membrane of cells. The Wzy protein was not well-expressed from a multi-copy plasmid in , or in K-12.

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1999-09-01
2019-12-13
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