Abstract
The flagellin gene was isolated from a Pseudomonas aeruginosa PAO1 genomic bank by conjugation into a PA103 Fla- strain. Flagellin DNA was transferred from motile recipient PA103 Fla+ cells by transformation into Escherichia coli. We show that transformed E. coli expresses flagellin protein. Export of flagellin to the E. coli cell surface was suggested by positive colony blots of unlysed cells and by isolation of flagellin protein from E. coli supernatants.
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- Allison J. S., Dawson M., Drake D., Montie T. C. Electrophoretic separation and molecular weight characterization of Pseudomonas aeruginosa H-antigen flagellins. Infect Immun. 1985 Sep;49(3):770–774. doi: 10.1128/iai.49.3.770-774.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ansorg R. Flagellaspezifisches H-Antigenschema von Pseudomonas aeruginosa. Zentralbl Bakteriol Orig A. 1978 Nov;242(2):228–238. [PubMed] [Google Scholar]
- Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Drake D., Montie T. C. Protection against Pseudomonas aeruginosa infection by passive transfer of anti-flagellar serum. Can J Microbiol. 1987 Sep;33(9):755–763. doi: 10.1139/m87-130. [DOI] [PubMed] [Google Scholar]
- Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finlay B. B., Pasloske B. L., Paranchych W. Expression of the Pseudomonas aeruginosa PAK pilin gene in Escherichia coli. J Bacteriol. 1986 Feb;165(2):625–630. doi: 10.1128/jb.165.2.625-630.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Flynn J. L., Ohman D. E. Cloning of genes from mucoid Pseudomonas aeruginosa which control spontaneous conversion to the alginate production phenotype. J Bacteriol. 1988 Apr;170(4):1452–1460. doi: 10.1128/jb.170.4.1452-1460.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gill P. R., Agabian N. A comparative structural analysis of the flagellin monomers of Caulobacter crescentus indicates that these proteins are encoded by two genes. J Bacteriol. 1982 May;150(2):925–933. doi: 10.1128/jb.150.2.925-933.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goldberg J. B., Ohman D. E. Cloning and expression in Pseudomonas aeruginosa of a gene involved in the production of alginate. J Bacteriol. 1984 Jun;158(3):1115–1121. doi: 10.1128/jb.158.3.1115-1121.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gray G. L., Smith D. H., Baldridge J. S., Harkins R. N., Vasil M. L., Chen E. Y., Heyneker H. L. Cloning, nucleotide sequence, and expression in Escherichia coli of the exotoxin A structural gene of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 1984 May;81(9):2645–2649. doi: 10.1073/pnas.81.9.2645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hedstrom R. C., Funk C. R., Kaper J. B., Pavlovskis O. R., Galloway D. R. Cloning of a gene involved in regulation of exotoxin A expression in Pseudomonas aeruginosa. Infect Immun. 1986 Jan;51(1):37–42. doi: 10.1128/iai.51.1.37-42.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hindahl M. S., Frank D. W., Iglewski B. H. Molecular studies of a positive regulator of toxin A synthesis in Pseudomonas aeruginosa. Antibiot Chemother (1971) 1987;39:279–289. doi: 10.1159/000414353. [DOI] [PubMed] [Google Scholar]
- Holder I. A., Wheeler R., Montie T. C. Flagellar preparations from Pseudomonas aeruginosa: animal protection studies. Infect Immun. 1982 Jan;35(1):276–280. doi: 10.1128/iai.35.1.276-280.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Homma M., Fujita H., Yamaguchi S., Iino T. Regions of Salmonella typhimurium flagellin essential for its polymerization and excretion. J Bacteriol. 1987 Jan;169(1):291–296. doi: 10.1128/jb.169.1.291-296.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Komeda Y., Kutsukake K., Iino T. Definition of additional flagellar genes in Escherichia coli K12. Genetics. 1980 Feb;94(2):277–290. doi: 10.1093/genetics/94.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kutsukake K., Iino T., Komeda Y., Yamaguchi S. Functional homology of fla genes between Salmonella typhimurium and Escherichia coli. Mol Gen Genet. 1980 Apr;178(1):59–67. doi: 10.1007/BF00267213. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Liu P. V. The roles of various fractions of Pseudomonas aeruginosa in its pathogenesis. 3. Identity of the lethal toxins produced in vitro and in vivo. J Infect Dis. 1966 Oct;116(4):481–489. doi: 10.1093/infdis/116.4.481. [DOI] [PubMed] [Google Scholar]
- Lory S., Strom M. S., Johnson K. Expression and secretion of the cloned Pseudomonas aeruginosa exotoxin A by Escherichia coli. J Bacteriol. 1988 Feb;170(2):714–719. doi: 10.1128/jb.170.2.714-719.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montie T. C., Anderson T. R. Enzyme-linked immunosorbent assay for detection of Pseudomonas aeruginosa H (flagellar) antigen. Eur J Clin Microbiol Infect Dis. 1988 Apr;7(2):256–260. doi: 10.1007/BF01963097. [DOI] [PubMed] [Google Scholar]
- Montie T. C., Doyle-Huntzinger D., Craven R. C., Holder I. A. Loss of virulence associated with absence of flagellum in an isogenic mutant of Pseudomonas aeruginosa in the burned-mouse model. Infect Immun. 1982 Dec;38(3):1296–1298. doi: 10.1128/iai.38.3.1296-1298.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montie T. C., Drake D., Sellin H., Slater O., Edmonds S. Motility, virulence, and protection with a flagella vaccine against Pseudomonas aeruginosa infection. Antibiot Chemother (1971) 1987;39:233–248. doi: 10.1159/000414349. [DOI] [PubMed] [Google Scholar]
- Nicas T. I., Frank D. W., Stenzel P., Lile J. D., Iglewski B. H. Role of exoenzyme S in chronic Pseudomonas aeruginosa lung infections. Eur J Clin Microbiol. 1985 Apr;4(2):175–179. doi: 10.1007/BF02013593. [DOI] [PubMed] [Google Scholar]
- Ogle J. W., Janda J. M., Woods D. E., Vasil M. L. Characterization and use of a DNA probe as an epidemiological marker for Pseudomonas aeruginosa. J Infect Dis. 1987 Jan;155(1):119–126. doi: 10.1093/infdis/155.1.119. [DOI] [PubMed] [Google Scholar]
- Silverman M., Simon M. I. Bacterial flagella. Annu Rev Microbiol. 1977;31:397–419. doi: 10.1146/annurev.mi.31.100177.002145. [DOI] [PubMed] [Google Scholar]
- Strom M. S., Lory S. Cloning and expression of the pilin gene of Pseudomonas aeruginosa PAK in Escherichia coli. J Bacteriol. 1986 Feb;165(2):367–372. doi: 10.1128/jb.165.2.367-372.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsuda M., Iino T. Ordering of the flagellar genes in Pseudomonas aeruginosa by insertions of mercury transposon Tn501. J Bacteriol. 1983 Feb;153(2):1008–1017. doi: 10.1128/jb.153.2.1008-1017.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsuda M., Iino T. Transductional analysis of the flagellar genes in Pseudomonas aeruginosa. J Bacteriol. 1983 Feb;153(2):1018–1026. doi: 10.1128/jb.153.2.1018-1026.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vasil M. L. Pseudomonas aeruginosa: biology, mechanisms of virulence, epidemiology. J Pediatr. 1986 May;108(5 Pt 2):800–805. doi: 10.1016/s0022-3476(86)80748-x. [DOI] [PubMed] [Google Scholar]