Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Jan 1991; 173(2): 505–513.
PMCID: PMC207039

Isolation and biochemical and molecular analyses of a species-specific protein antigen from the gastric pathogen Helicobacter pylori.


A protein of Mr 26,000 which was present in large quantities in extracts of cells of Helicobacter pylori was purified to homogeneity by ammonium sulfate precipitation followed by gel filtration and reversed-phase chromatography or anion-exchange chromatography. The protein appeared to be associated with the soluble fraction of the cell, and antibodies raised against the protein were reactive with whole-cell lysates of a variety of H. pylori strains in a simple immunodot blot assay. This reaction was species specific. Protein sequence determination of the amino terminus and internal cyanogen bromide fragments and amino acid composition analysis were performed. An oligonucleotide derived from these data was used to clone a fragment encoding most of the coding sequence. Expression in Escherichia coli was dependent on vector promoters. The DNA sequence of the fragment was determined. DNA probes derived from the cloned fragment hybridized to genomic DNA of all H. pylori strains tested, but not to DNAs of Helicobacter mustelae, Wolinella succinogenes, various Campylobacter species, and a panel of gram-negative enteric bacteria. The apparent uniqueness of this protein may be exploited for the development of species-specific diagnostics for this gastric pathogen.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Amann E, Brosius J. "ATG vectors' for regulated high-level expression of cloned genes in Escherichia coli. Gene. 1985;40(2-3):183–190. [PubMed]
  • Belland RJ, Trust TJ. Deoxyribonucleic acid sequence relatedness between thermophilic members of the genus Campylobacter. J Gen Microbiol. 1982 Nov;128(11):2515–2522. [PubMed]
  • Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich EC. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. [PubMed]
  • Blaser MJ. Gastric Campylobacter-like organisms, gastritis, and peptic ulcer disease. Gastroenterology. 1987 Aug;93(2):371–383. [PubMed]
  • Buck GE, Gourley WK, Lee WK, Subramanyam K, Latimer JM, DiNuzzo AR. Relation of Campylobacter pyloridis to gastritis and peptic ulcer. J Infect Dis. 1986 Apr;153(4):664–669. [PubMed]
  • Clayton CL, Wren BW, Mullany P, Topping A, Tabaqchali S. Molecular cloning and expression of Campylobacter pylori species-specific antigens in Escherichia coli K-12. Infect Immun. 1989 Feb;57(2):623–629. [PMC free article] [PubMed]
  • Dooley CP, Cohen H. The clinical significance of Campylobacter pylori. Ann Intern Med. 1988 Jan;108(1):70–79. [PubMed]
  • Evans DG, Evans DJ, Jr, Moulds JJ, Graham DY. N-acetylneuraminyllactose-binding fibrillar hemagglutinin of Campylobacter pylori: a putative colonization factor antigen. Infect Immun. 1988 Nov;56(11):2896–2906. [PMC free article] [PubMed]
  • Evans DJ, Jr, Evans DG, Graham DY, Klein PD. A sensitive and specific serologic test for detection of Campylobacter pylori infection. Gastroenterology. 1989 Apr;96(4):1004–1008. [PubMed]
  • Filip C, Fletcher G, Wulff JL, Earhart CF. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. [PMC free article] [PubMed]
  • Graham DY, Klein PD. Campylobacter pyloridis gastritis: the past, the present, and speculations about the future. Am J Gastroenterol. 1987 Apr;82(4):283–286. [PubMed]
  • Graham DY, Klein PD, Opekun AR, Boutton TW. Effect of age on the frequency of active Campylobacter pylori infection diagnosed by the [13C]urea breath test in normal subjects and patients with peptic ulcer disease. J Infect Dis. 1988 Apr;157(4):777–780. [PubMed]
  • Grunstein M, Hogness DS. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. [PMC free article] [PubMed]
  • Hawley DK, McClure WR. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. [PMC free article] [PubMed]
  • Hunkapiller MW, Hood LE. Analysis of phenylthiohydantoins by ultrasensitive gradient high-performance liquid chromatography. Methods Enzymol. 1983;91:486–493. [PubMed]
  • Jiang SJ, Liu WZ, Zhang DZ, Shi Y, Xiao SD, Zhang ZH, Lu DY. Campylobacter-like organisms in chronic gastritis, peptic ulcer, and gastric carcinoma. Scand J Gastroenterol. 1987 Jun;22(5):553–558. [PubMed]
  • Krakowka S, Morgan DR, Kraft WG, Leunk RD. Establishment of gastric Campylobacter pylori infection in the neonatal gnotobiotic piglet. Infect Immun. 1987 Nov;55(11):2789–2796. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lambert JR, Borromeo M, Pinkard KJ, Turner H, Chapman CB, Smith ML. Colonization of gnotobiotic piglets with Campylobacter Pyloridis--an animal model? J Infect Dis. 1987 Jun;155(6):1344–1344. [PubMed]
  • Lior H, Woodward DL, Edgar JA, Laroche LJ, Gill P. Serotyping of Campylobacter jejuni by slide agglutination based on heat-labile antigenic factors. J Clin Microbiol. 1982 May;15(5):761–768. [PMC free article] [PubMed]
  • Lipman DJ, Pearson WR. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. [PubMed]
  • Logan SM, Trust TJ. Molecular identification of surface protein antigens of Campylobacter jejuni. Infect Immun. 1983 Nov;42(2):675–682. [PMC free article] [PubMed]
  • Logan SM, Trust TJ. Location of epitopes on Campylobacter jejuni flagella. J Bacteriol. 1986 Nov;168(2):739–745. [PMC free article] [PubMed]
  • Logan SM, Trust TJ, Guerry P. Evidence for posttranslational modification and gene duplication of Campylobacter flagellin. J Bacteriol. 1989 Jun;171(6):3031–3038. [PMC free article] [PubMed]
  • Majewski SI, Goodwin CS. Restriction endonuclease analysis of the genome of Campylobacter pylori with a rapid extraction method: evidence for considerable genomic variation. J Infect Dis. 1988 Mar;157(3):465–471. [PubMed]
  • Marshall BJ, Armstrong JA, McGechie DB, Glancy RJ. Attempt to fulfil Koch's postulates for pyloric Campylobacter. Med J Aust. 1985 Apr 15;142(8):436–439. [PubMed]
  • Marshall BJ, Surveyor I. Carbon-14 urea breath test for the diagnosis of Campylobacter pylori associated gastritis. J Nucl Med. 1988 Jan;29(1):11–16. [PubMed]
  • McCoy EC, Doyle D, Burda K, Corbeil LB, Winter AJ. Superficial antigens of Campylobacter (Vibrio) fetus: characterization of antiphagocytic component. Infect Immun. 1975 Mar;11(3):517–525. [PMC free article] [PubMed]
  • Misra TK. A new strategy to create ordered deletions for rapid nucleotide sequencing. Gene. 1985;34(2-3):263–268. [PubMed]
  • Mobley HL, Hausinger RP. Microbial ureases: significance, regulation, and molecular characterization. Microbiol Rev. 1989 Mar;53(1):85–108. [PMC free article] [PubMed]
  • Morotomi M, Hoshina S, Green P, Neu HC, LoGerfo P, Watanabe I, Mutai M, Weinstein IB. Oligonucleotide probe for detection and identification of Campylobacter pylori. J Clin Microbiol. 1989 Dec;27(12):2652–2655. [PMC free article] [PubMed]
  • Morris A, Nicholson G. Ingestion of Campylobacter pyloridis causes gastritis and raised fasting gastric pH. Am J Gastroenterol. 1987 Mar;82(3):192–199. [PubMed]
  • Owen RJ. Nucleic acids in the classification of Campylobacters. Eur J Clin Microbiol. 1983 Aug;2(4):367–377. [PubMed]
  • Pridmore RD. New and versatile cloning vectors with kanamycin-resistance marker. Gene. 1987;56(2-3):309–312. [PubMed]
  • Robertson EF, Dannelly HK, Malloy PJ, Reeves HC. Rapid isoelectric focusing in a vertical polyacrylamide minigel system. Anal Biochem. 1987 Dec;167(2):290–294. [PubMed]
  • Romaniuk PJ, Zoltowska B, Trust TJ, Lane DJ, Olsen GJ, Pace NR, Stahl DA. Campylobacter pylori, the spiral bacterium associated with human gastritis, is not a true Campylobacter sp. J Bacteriol. 1987 May;169(5):2137–2141. [PMC free article] [PubMed]
  • Simor AE, Shames B, Drumm B, Sherman P, Low DE, Penner JL. Typing of Campylobacter pylori by bacterial DNA restriction endonuclease analysis and determination of plasmid profile. J Clin Microbiol. 1990 Jan;28(1):83–86. [PMC free article] [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet. 1983 Jun 4;1(8336):1273–1275. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...