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Appl Environ Microbiol. Dec 1997; 63(12): 4866–4871.
PMCID: PMC168813

Construction of a Helicobacter pylori-Escherichia coli shuttle vector for gene transfer in Helicobacter pylori.


In this study, a Helicobacter pylori-Escherichia coli shuttle vector was constructed for transferring DNA into H. pylori. The smallest cryptic plasmid (1.2 kb), pHP489, among those harbored by 77 H. pylori isolates was selected as a base replicon for constructing vectors. HindIII-digested pHP489 was ligated with a kanamycin resistance gene [aph(3')-III], which originated from Campylobacter jejuni, to produce the recombinant plasmid pHP489K. pHP489K was efficiently transformed into and stably maintained in H. pylori strains. The shuttle vector pBHP489K (3.6 kb) was constructed by the recombination of pHP489, ColE1, and aph(3')-III sequences. pBHP489K was reciprocally transformed into and maintained in both H. pylori and E. coli. Introduction of the shuttle vector clone DNA (pBHP489K/AB; 6.7 kb), containing the ureA and ureB genes of H. pylori, into urease-negative mutants of H. pylori led to the restoration of their urease activity. The transformants were confirmed to contain the incoming plasmid DNA. pBHP489K satisfied the requirements for an H. pylori-E. coli shuttle vector, implying that it might be a useful vector for investigating pathogenicity and restriction-modification systems of H. pylori.

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  • Baik SC, Youn HS, Chung MH, Lee WK, Cho MJ, Ko GH, Park CK, Kasai H, Rhee KH. Increased oxidative DNA damage in Helicobacter pylori-infected human gastric mucosa. Cancer Res. 1996 Mar 15;56(6):1279–1282. [PubMed]
  • Bukanov NO, Berg DE. Ordered cosmid library and high-resolution physical-genetic map of Helicobacter pylori strain NCTC11638. Mol Microbiol. 1994 Feb;11(3):509–523. [PubMed]
  • Covacci A, Censini S, Bugnoli M, Petracca R, Burroni D, Macchia G, Massone A, Papini E, Xiang Z, Figura N, et al. Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5791–5795. [PMC free article] [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Ferrero RL, Cussac V, Courcoux P, Labigne A. Construction of isogenic urease-negative mutants of Helicobacter pylori by allelic exchange. J Bacteriol. 1992 Jul;174(13):4212–4217. [PMC free article] [PubMed]
  • Graham DY. Campylobacter pylori and peptic ulcer disease. Gastroenterology. 1989 Feb;96(2 Pt 2 Suppl):615–625. [PubMed]
  • Gruss A, Ehrlich SD. The family of highly interrelated single-stranded deoxyribonucleic acid plasmids. Microbiol Rev. 1989 Jun;53(2):231–241. [PMC free article] [PubMed]
  • Heuermann D, Haas R. Genetic organization of a small cryptic plasmid of Helicobacter pylori. Gene. 1995 Nov 7;165(1):17–24. [PubMed]
  • Kaldor J, Tee W, Nicolacopolous C, Demirtzoglou K, Noonan D, Dwyer B. Immunoblot confirmation of immune response to Campylobacter pyloridis in patients with duodenal ulcers. Med J Aust. 1986 Aug 4;145(3-4):133–135. [PubMed]
  • Kleanthous H, Clayton CL, Tabaqchali S. Characterization of a plasmid from Helicobacter pylori encoding a replication protein common to plasmids in gram-positive bacteria. Mol Microbiol. 1991 Oct;5(10):2377–2389. [PubMed]
  • Labigne A, Cussac V, Courcoux P. Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity. J Bacteriol. 1991 Mar;173(6):1920–1931. [PMC free article] [PubMed]
  • Labigne-Roussel A, Harel J, Tompkins L. Gene transfer from Escherichia coli to Campylobacter species: development of shuttle vectors for genetic analysis of Campylobacter jejuni. J Bacteriol. 1987 Nov;169(11):5320–5323. [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]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet. 1983 Jun 4;1(8336):1273–1275. [PubMed]
  • Minnis JA, Taylor TE, Knesek JE, Peterson WL, McIntire SA. Characterization of a 3.5-kbp plasmid from Helicobacter pylori. Plasmid. 1995 Jul;34(1):22–36. [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]
  • Nedenskov-Sørensen P, Bukholm G, Bøvre K. Natural competence for genetic transformation in Campylobacter pylori. J Infect Dis. 1990 Feb;161(2):365–366. [PubMed]
  • Nomura A, Stemmermann GN, Chyou PH, Kato I, Perez-Perez GI, Blaser MJ. Helicobacter pylori infection and gastric carcinoma among Japanese Americans in Hawaii. N Engl J Med. 1991 Oct 17;325(16):1132–1136. [PubMed]
  • Olivieri R, Bugnoli M, Armellini D, Bianciardi S, Rappuoli R, Bayeli PF, Abate L, Esposito E, de Gregorio L, Aziz J, et al. Growth of Helicobacter pylori in media containing cyclodextrins. J Clin Microbiol. 1993 Jan;31(1):160–162. [PMC free article] [PubMed]
  • Park IS, Hausinger RP. Requirement of carbon dioxide for in vitro assembly of the urease nickel metallocenter. Science. 1995 Feb 24;267(5201):1156–1158. [PubMed]
  • Parsonnet J, Hansen S, Rodriguez L, Gelb AB, Warnke RA, Jellum E, Orentreich N, Vogelman JH, Friedman GD. Helicobacter pylori infection and gastric lymphoma. N Engl J Med. 1994 May 5;330(18):1267–1271. [PubMed]
  • Penfold SS, Lastovica AJ, Elisha BG. Demonstration of plasmids in Campylobacter pylori. J Infect Dis. 1988 Apr;157(4):850–851. [PubMed]
  • Phadnis SH, Westblom TU, Normark S. Molecular cloning of Helicobacter pylori DNA: important differences between mcrBC deletion host strains. Mol Microbiol. 1993 Dec;10(5):1151–1151. [PubMed]
  • Schmitt W, Odenbreit S, Heuermann D, Haas R. Cloning of the Helicobacter pylori recA gene and functional characterization of its product. Mol Gen Genet. 1995 Sep 20;248(5):563–572. [PubMed]
  • Segal ED, Shon J, Tompkins LS. Characterization of Helicobacter pylori urease mutants. Infect Immun. 1992 May;60(5):1883–1889. [PMC free article] [PubMed]
  • Shizuya H, Birren B, Kim UJ, Mancino V, Slepak T, Tachiiri Y, Simon M. Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8794–8797. [PMC free article] [PubMed]
  • Taylor DE. Genetics of Campylobacter and Helicobacter. Annu Rev Microbiol. 1992;46:35–64. [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]
  • Tsuda M, Karita M, Nakazawa T. Genetic transformation in Helicobacter pylori. Microbiol Immunol. 1993;37(1):85–89. [PubMed]
  • Wang Y, Taylor DE. Natural transformation in Campylobacter species. J Bacteriol. 1990 Feb;172(2):949–955. [PMC free article] [PubMed]
  • Wang Y, Roos KP, Taylor DE. Transformation of Helicobacter pylori by chromosomal metronidazole resistance and by a plasmid with a selectable chloramphenicol resistance marker. J Gen Microbiol. 1993 Oct;139(10):2485–2493. [PubMed]
  • Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet. 1983 Jun 4;1(8336):1273–1275. [PubMed]

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