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Stable surface expression of a gene for Helicobacter pylori toxic porin protein with pBAD expression system

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Summary

Helicobacter pylori (H. pylori) infection causes peptic and duodenal diseases in humans. Among a 32-protein family of outer membrane proteins, a porin-like protein, HopE, has been a subject of note, mainly for its conservative nature among H. pylori, and for its potential as a vaccine candidate. To achieve stable surface expression of this host cell-toxic protein, hopE gene was introduced into pBAD expression system. After induction with arabinose, all 15 randomly-chosen E. coli LMG 194 colonies from 3 successive passages could express HopE protein, while only 1 from 5 E. coli colonies that contained lac operon-regulated plasmid encoding hopE gene could express HopE. Indirect immunofluorescence confirmed the expression of HopE on E. coli cell surface.

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References

  1. Snaith A, El-Omar EM. Helicobacter pylori: host genetics and disease outcomes. Expert Rev Gastroenterol Hepatol, 2008, 2(4):577–585

    Article  PubMed  CAS  Google Scholar 

  2. Clayton CL, Harry LT. Mobley. Helicobacter pylori Protocols. Humana Press: New Jersey. 1997.58

    Book  Google Scholar 

  3. Go M. Natural history and epidemiology of Helicobacter pylori infection. Aliment Pharmacol Ther, 2002,16(Suppl):3–15

    Article  PubMed  Google Scholar 

  4. Silva DG, Stevens RH, Macedo JM, et al. Detection of cytotoxin genotypes of Helicobacter pylori in stomach, saliva and dental plaque. Arch Oral Biol, 2009,54(7): 684–688

    Article  PubMed  CAS  Google Scholar 

  5. Alm RA, Ling LS, Moir DT, et al. Genomic-sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori. Nature, 1999,397(6715): 176–180

    Article  PubMed  Google Scholar 

  6. Tomb JF, White O, Kerlavage AR, et al. The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature, 1997,388(6642):539–547

    Article  PubMed  CAS  Google Scholar 

  7. Exner MM, Doig P, Trust TJ, et al. Isolation and characterization of a family of porin proteins from Helicobacter pylori. Infect Immun, 1995,63(4):1567–1572

    PubMed  CAS  Google Scholar 

  8. Doig P, Exner MM, Hancock RE, et al. Isolation and characterization of a conserved porin protein from Helicobacter pylori. J Bacteriol, 1995,177(19):5447–5452

    PubMed  CAS  Google Scholar 

  9. Tufano MA, Rossano F, Catalanotti P, et al. Immunobiological activities of Helicobacter pylori porins. Infect Immun, 1994,62(4):1392–1399

    PubMed  CAS  Google Scholar 

  10. Pullen JK, Liang SM, Blake MS, et al. Production of Haemophilus influenzae type-b porin in Escherichia coli and its folding into the trimeric form. Gene, 1995,152: 85–88

    Article  PubMed  CAS  Google Scholar 

  11. Schmid B, Krömer M, Schulz G E. Expression of porin from Rhodopseudomonas blastica in Escherichia coli inclusion bodies and folding into exact native structure. FEBS Lett, 1996,381(1–2):111–114

    Article  PubMed  CAS  Google Scholar 

  12. Senaratne RH, Mobasheri H, Papavinasasundaram KG, et al. Expression of a gene for a porin-like protein of the OmpA family from Mycobacterium tuberculosis H37Rv. J Bacteriol, 1998,180(14):3541–3547

    PubMed  CAS  Google Scholar 

  13. Guzman LM, Belin D, Carson MJ, et al. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol, 1995, 177(14):4121–4130

    PubMed  CAS  Google Scholar 

  14. Altschul SF, Gish W, Miller W, et al. Basic local alignment search tool. J Mol Biol, 1990,215(3):403–410

    PubMed  CAS  Google Scholar 

  15. Bina J, Bains M, Hancock RE. Functional expression in Escherichia coli and membrane topology of porin HopE, a member of a large family of conserved proteins in Helicobacter pylori. J Bacteriol, 2000,182(9):2370–2375

    Article  PubMed  CAS  Google Scholar 

  16. Labigne A, Cussac V, Courcoux P. Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity. J Bacteriol, 1991,173(6): 1920–1931

    PubMed  CAS  Google Scholar 

  17. Qiu D, Damron FH, Mima T, et al. pBAD-based shuttle vectors for functional analysis of toxic and highly regulated genes in Pseudomonas and Burkholderia spp. and other bacteria. Appl Environ Microbiol, 2008,74(23): 7422–7426

    Article  PubMed  CAS  Google Scholar 

  18. Mardanov AV, Strakhova TS, Smagin VA, et al. Tightly regulated, high-level expression from controlled copy number vectors based on the replicon of temperate phage N15. Gene, 2007,395(1–2):15–21

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Endodontics, Guanghua College of Stomatology, Sun Yat-Sen University, Guangzhou, 510055, China

    Zhixiang Peng  (彭志翔), Xi Wei  (韦 曦) & Zhengmei Lin  (林正梅)

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  1. Zhixiang Peng  (彭志翔)
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  2. Xi Wei  (韦 曦)
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  3. Zhengmei Lin  (林正梅)
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Peng, Z., Wei, X. & Lin, Z. Stable surface expression of a gene for Helicobacter pylori toxic porin protein with pBAD expression system. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 29, 435–438 (2009). https://doi.org/10.1007/s11596-009-0409-5

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  • DOI: https://doi.org/10.1007/s11596-009-0409-5

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