Non‐motile mutants of Helicobacter pylori and Helicobacter mustelae defective in flagellar hook production
- 1 November 1994
- journal article
- Published by Wiley in Molecular Microbiology
- Vol. 14 (4) , 691-703
- https://doi.org/10.1111/j.1365-2958.1994.tb01307.x
Abstract
Flagellar hooks were purified from Helicobacter pylori and Helicobacter mustelae. The 70 × 16nm H. pylori hook was composed of FIgE subunits of 78kDa, while the 72 × 16nm H. mustelae hook was composed of 87kDa subunits. N-terminal sequence was obtained for the FIgH proteins of both species, and for an internal H. mustelae FlgE peptide. Degenerate oligonucleotide primers allowed amplification of a 1.2 kb fragment from the H. mustelae chromosome, which carried part of the flgE gene. The corresponding H. pylori gene was cloned by immunoscreening of a genomic library constructed in λZAP Express, The translated H. pylori flgE sequence indicated a protein with limited homology with the hook proteins from Salmonella typhimurium and Treponema phagedenis. Mutants of H. pylori and H. mustelae defective in hook production generated by allele replacement were non-motile and devoid of flagellar filaments but produced both flagellin subunits, which were localized in the soluble fraction of the cell. The level of flagellin production was unchanged in the mutants, indicating that the regulation of flagellin expression in Helicobacter differs from that in the Enterobacteriaceae.Keywords
This publication has 53 references indexed in Scilit:
- Significance of Duplicated Flagellin Genes in CampylobacterJournal of Molecular Biology, 1993
- Organization and ordered expression of Caulobacter genes encoding flagellar basal body rod and ring proteinsJournal of Molecular Biology, 1992
- Subdivision of flagellar region III of the Escherichia coli and Salmonella typhimurium chromosomes and identification of two additional flagellar genesJournal of General Microbiology, 1992
- Flagellar hook and hook-associated proteins of Salmonella typhimurium and their relationship to other axial components of the flagellumJournal of Molecular Biology, 1990
- Rapid extraction of bacterial genomic DNA with guanidium thiocyanateLetters in Applied Microbiology, 1989
- New and versatile cloning vectors with kanamycin-resistance markerGene, 1987
- Transcriptional regulation of a periodically controlled flagellar gene operon in Caulobacter crescentusJournal of Molecular Biology, 1985
- Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectorsGene, 1985
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- A general method applicable to the search for similarities in the amino acid sequence of two proteinsJournal of Molecular Biology, 1970