1887

Microbiology Society logo

Volume 143, Issue 11

Expression of lipo-oligosaccharide (LOS) antigens in Free

Abstract

spp. are well recognized as primary pathogens in animals and in people. To isolate and define the genetic regions encoding major surface antigens of genomic DNA of the type strain of the species, RMIT-32A, was cloned into a cosmid vector, pLA2917, in and the resulting genomic library was screened using antiserum raised to the parent strain. Six cosmid clones were found to express a series of immunoreactive bands in the 15-25 kDa range. These bands were proteinase K-resistant and were found in the LPS fraction of the cells, suggesting that the recombinant cosmids expressed lipo-oligosaccharide (LOS) antigen(). The minimum DNA insert size required for expression of LOS antigen() in was 11-8 kb. This region was subcloned into the plasmid vector pBR322. The partial sequencing of the 11.8 kb region showed that it contains two ORFs, designated and showing homology with the gene from and the gene from Both genes are involved in LPS synthesis. The region also contained a sequence homologous to the gene of and which is involved in core oligosaccharide synthesis.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Campylobacter , cloning and expression , lipo-oligosaccharide (LOS) and LPS
© Society for General Microbiology 1997
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-11-3481
1997-11-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/11/mic-143-11-3481.html?itemId=/content/journal/micro/10.1099/00221287-143-11-3481&mimeType=html&fmt=ahah

References

  1. Alderton M.R., Borland R., Coloe P.J. 1991; Experimental reproduction of porcine proliferative enteritis.. J Comp Pathol 106:159–167
     [Google Scholar]
  2. Alderton M.R., Korolik V., Coloe P.J., Dewhirst F.E., Paster B.J. 1995; ‘Campylobacter hyoilei’ sp. nov.: associated with porcine proliferative enteritis.. Int J Syst Bacteriol 45:61–66
     [Google Scholar]
  3. Allen A., Maskell D.J. 1995; Identification, cloning and mutagenesis of a genetic locus required for lipopolysaccharide biosynthesis in Bordetella pertussis.. Mol Microbiol 19:19–37
     [Google Scholar]
  4. Allen L.N., Hanson R.S. 1985; Construction of broad-host-range cosmid cloning vectors: identification of genes necessary for growth of Methylobacterium organophilum on methanol.. J Bacteriol 161:955–962
     [Google Scholar]
  5. Altschul A.F., Gish W., Miller W., Myers E.W., Lipman D.J. 1990; Basic local alignment search tool.. J Mol Biol 215:403–410
     [Google Scholar]
  6. Arakawa Y., Ohta M., Wacharotayankun R., Mori M., Kido N., Ito H., Komatsu T., Sugiyama T., Kato N. 1991; Biosynthesis of Klebsiella K2 capsular polysaccharide in Escherichia coli HB101 requires the functions of rmpA and the chromosomal cps gene cluster of the virulent strain Klebsiella pneumoniae Chedid (Ol :K2).. Infect Immun 59:2043–2050
     [Google Scholar]
  7. Arakawa Y., Wacharotayankun R., Nagatsuka T., Ito H., Kato N., Ohta M. 1995; Genomic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis in the virulent strain Chedid.. J Bacteriol 177:1788–1796
     [Google Scholar]
  8. Aspinall G.O., McDonald A.G., Raju T.S., Pang H., Mills S.D., Kurjanczyk L.A., Penner J.L. 1992; Serological diversity and chemical structures of Campylobacter jejuni low molecular weight lipopolysaccharides.. J Bacteriol 174:1324–1332
     [Google Scholar]
  9. Aspinall G.O., McDonald A.G., Pang H., Kurjanczyk L.A., Penner J.L. 1993a; Lipopolysaccharide of Campylobacter coli serotype 0:30. Fractionation and structure of liberated core oligosaccharide.. J Biol Chem 268:6263–6268
     [Google Scholar]
  10. Aspinall G.O., McDonald A.G., Raju T.S., Pang H., Kurjanczyk L.A., Penner J.L., Moran A.P. 1993b; Chemical structure of the core region of Campylobacter jejuni serotype O:2 lipopolysaccharide.. Eur J Biochem 213:1029–1037
     [Google Scholar]
  11. Aspinall G.O., McDonald A.G., Raju T.S., Pang H., Moran A.P., Penner J.L. 1993c; Chemical structures of the core regions of Campylobacter jejuni serotypes 0:1, 0:4, 0:23, and 0:36 lipopolysaccharides.. Eur J Biochem 216:880
     [Google Scholar]
  12. Aspinall G.O., Fujimoto S., McDonald A.G., Pang H., Kurjanczyk L.A., Penner J.L. 1994a; Lipopolysaccharides from Campylobacter jejuni associated with Guillain-Barré syndrome patients mimic human gangliosides in structure.. Infect Immun 62:2122–2125
     [Google Scholar]
  13. Aspinall G.O., McDonald A.G., Pang H., Kurjanczyk L.A., Penner J.L. 1994b; Lipopolysaccharides of Campylobacter jejuni serotype 0:19: structures of core oligosaccharide regions from the serostrain and two bacterial isolates from patients with the Guillain-Barre syndrome.. Biochemistry 33:241–249
     [Google Scholar]
  14. Aspinall G.O., Lynch C.M., Pang H., Shaver R.T., Moran A.P. 1995a; Chemical structures of the core region of Campylobacter jejuni 0:3 lipopolysaccharide and an associated polysaccharide.. Eur J Biochem 231:570–578
     [Google Scholar]
  15. Aspinall G.O., Monteiro M.A., Pang H. 1995b; Lipo-oligosaccharide of the Campylobacter lari type strain ATCC 35221. Structure of the liberated oligosaccharide and an associated extracellular polysaccharide.. Carbohydr Res 279:245–264
     [Google Scholar]
  16. Aspinall G.O., Monteiro M.A., Pang H., Kurjanczyk L.A., Penner J.L. 1995c; Lipo-oligosaccharide of Campylobacter lari strain PC 637. Structure of the liberated oligosaccharide and an associated extracellular polysaccharide.. Carbohydr Res 279:227–244
     [Google Scholar]
  17. Bachmann B.J. 1987; Linkage map of Escherichia coli K12, edition 7.. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology pp. 807–876 Edited by Neidhardt F. C., Ingraham J. L., Brooks Low K., Magasanik B., Schaechter M., Umbarger H. E. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  18. Blaser M.J., Reller L.B. 1981; Campylobacter enteritis.. N Engl J Med 305:1444–1452
     [Google Scholar]
  19. Bolivar F., Rodriguez R.C., Greene P.J., Betlach M.C., Hey-necker H.C, Boyer H. W., Crosa J.H., Falkow S. 1977; Construction and characterisation of new cloning vehicles. II. A multipurpose cloning system.. Gene 2:95–113
     [Google Scholar]
  20. Bugert P., Geider K. 1995; Molecular analysis of ams operon required for exopolysaccharide synthesis of Erwinia amylovora.. Mol Microbiol 15:917–933
     [Google Scholar]
  21. Chan V.L., Bingham M., Kibue A., Nayudu P.R.V., Penner J.L. 1988; Cloning and expression of the Campylobacter jejuni glyA gene in Escherichia coli.. Gene 73:185–191
     [Google Scholar]
  22. Chen L., Coleman W.G. Jr 1993; Cloning and characterization of the Escherichia coli K-12 rfa-2 (rfaC) gene, a gene required for lipopolysaccharide inne core synthesis.. J Bacteriol 175:2534–2540
     [Google Scholar]
  23. Collins D.M., Ross D.E. 1984; Restriction endonuclease analysis of Campylobacter strains with particular reference to Campylobacter fetus ss fetus.. J Med Microbiol 18:117–124
     [Google Scholar]
  24. Collins L.V., Hackett J. 1991; Molecular cloning, characterisation, and nucleotide sequence of the rfc gene, which encodes an O-antigen polymerase of Salmonella typhimurium.. J Bacteriol 173:2521–2529
     [Google Scholar]
  25. Demarco de Hormaeche R., Macpherson A., Bowe F., Hormaeche C.E. 1991; Alterations of the LPS determine virulence of Neisseria gonorrhoeae in guinea-pig subcutaneous chambers.. Microb Pathog 11:159–170
     [Google Scholar]
  26. Fleischmann R.D., Adams M.D., White O. 37 other authors 1995; Whole-genome sequencing and assembly of Haemophilus influenzae Rd.. Science 269:496–512
     [Google Scholar]
  27. Frey J., Bagdasarian M., Feiss D., Franklin F.C.H., Deshusses J. 1983; Stable cosmid vectors that enable the introduction of cloned fragments into a wide range of Gram-negative bacteria.. Gene 24:299–308
     [Google Scholar]
  28. Gray J.X., Djordjeviv M.A., Rolfe B.G. 1990; Two genes that regulate exopolysaccharide production in Rhizobium sp. strain NGR234: DNA sequences and resultant phenotypes.. J Bacteriol 172:193–203
     [Google Scholar]
  29. Jiang X.-M., Neal B., Santiago F., Lee S.J., Romana L.K., Reeves P.R. 1991; Structure and sequence of the rfb (O antigen) gene cluster of Salmonella serovar typhimurium (strain LT2).. Mol Microbiol 5:695–713
     [Google Scholar]
  30. Klena J.D., Ashford R.S. II Schnaitman C.A. 1992; Role of Escherichia coli K-12 rfa genes and the rfp gene of Shigella dysenteriae 1 in generation of lipopolysaccharide core heterogeneity and attachment of O antigen.. J Bacteriol 174:7297–7307
     [Google Scholar]
  31. Korn L.J., Queen C. 1984; Analysis of biological sequences on small computers.. DNA 3:421–436
     [Google Scholar]
  32. Korolik V., Coloe P.J., Krishnapillai V. 1988; A specific DNA probe for the identification of Campylobacter jejuni.. J Gen Microbiol 134:521–529
     [Google Scholar]
  33. Korolik V., Moorthy L., Coloe P.J. 1995; Differentiation of Campylobacter jejuni and Campylobacter coli strains using restriction endonuclease DNA cleavage profiles and DNA fragment polymorphisms.. J Clin Microbiol 33:1136–1140
     [Google Scholar]
  34. Kuan S.K.C., Coloe P.J., Alderton M.R. 1992; Production of a monoclonal antibody that recognises the lipopolysaccharide of a Campylobacter-like organism.. Microbiol Immunol 36:791–801
     [Google Scholar]
  35. Labigne-Roussel A., Harel J., Tompkins L. 1987; Gene transfer from Escherichia coli to Campylobacter species: development of shuttle vectors for genetic analysis of Campylobacter jejuni.. J Bacteriol 169:5320–5323
     [Google Scholar]
  36. Laemmli U.K. 1970; Change of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
     [Google Scholar]
  37. Lawrence C.E., Stephen F.A., Boguski M.S., Neuwald A.F., Wootton J.C. 1993; Detecting subtle sequence signals: a Gibbs sampling strategy for multiple alignment.. Science 262:208–214
     [Google Scholar]
  38. Luderitz O., Freudenberg M.A., Galano C., Lehmann V., Rietschel E.T., Show D.H. 1982; Lipopolysaccharides of Gram-negative bacteria.. Curr Top Membr Transp 17:79–151
     [Google Scholar]
  39. Mandatori R., Penner J. L. 1989; Structural and antigenic properties of Campylobacter coli lipopolysaccharides.. Infect Immun 573506–3511
     [Google Scholar]
  40. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  41. Manning P.A., Heuzenroeder M.W., Yeadon J., Leavesley D.I., Reeves P.R., Rowley D. 1988; Molecular cloning and expression in Escherichia coli K-12 of the O-antigens of the Inaba and Ogawa serotypes of the Vibrio cholerae Ol lipopoly-saccharides and their potential for vaccine development.. Infect Immun 53:272–277
     [Google Scholar]
  42. Moxon R.E., Maskell D. 1992; Haemophilus influenzae lipopolysaccharide: the biochemistry and biology of a virulence factor.. In Molecular Biology of Bacterial Infection: Current Status and Future Perspectives pp. 75–96 Edited by Hormaeche C. E., Penn C. W., Smyth C. J. Cambridge: Cambridge University Press;
     [Google Scholar]
  43. Penner J.L., Hennessy J.N. 1980; Passive hemagglutination technique for serotyping Campylobacter fetus subsp. jejuni on the basis of soluble heat-stable antigens.. J Clin Microbiol 12:732–737
     [Google Scholar]
  44. Perez-Perez G.I., Biaser M.J. 1985; Lipopolysaccharide characteristics of pathogenic Campylobacters.. Infect Immun 47:353–359
     [Google Scholar]
  45. Perez-Perez G.I., Hopkins J.A., Biaser M.J. 1986; Lipopolysaccharide structures in Enterobacteriaceae, Pseudomonas aerugi-nosa and Vibrio cholerae are immunologically related to Campylobacter spp.. Infect Immun 51:204–208
     [Google Scholar]
  46. Preston A., Mandrell R.E., Gibson B.W., Apicella M.A. 1996; The lipooligosaccharides of pathogenic Gram-negative bacteria.. Crit Rev Microbiol 22:139–180
     [Google Scholar]
  47. Reeves P. 1993; Evolution of Salmonella O antigen variation by interspecific gene transfer on a large scale.. Trends Genet 9:17–22
     [Google Scholar]
  48. Salloway S., Mermel L.A., Seamans M., Aspinall G.O., Nam Shin J.E., Kurjanczyk L.A., Penner J.L. 1996; Miller-Fisher syndrome associated with Campylobacter jejuni bearing lipopolysaccharide molecules that mimic human ganglioside GD3.. Infect Immun 64:2945–2949
     [Google Scholar]
  49. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  50. Sanderson K.E., Hurley J.A. 1987; Linkage map of Salmonella typhimurium.. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology pp. 877–918 Edited by Neidhardt F. C., Ingraham J. L., Brooks Low K., Magasanik B., Schaechter M., Umbarger H. E. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  51. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain-terminating inhibitors.. Proc Natl Acad Sci USA 745463–5467
     [Google Scholar]
  52. Schwerer B., Neisser A., Polt R.J., Bernheimer H., Moran A.P. 1995; Antibody cross-reactivities between gangliosides and lipopolysaccharides of Campylobacter jejuni serotypes associated with Guillain-Barré syndrome.. J Endotoxin Res 2:395–403
     [Google Scholar]
  53. Sirisena D.M., Brozek K.A., Maclachlan P.R., Sanderson K.E., Raetz C.R.H. 1992; The rfaC gene of Salmonella typhimurium.. J Biol Chem 267:18874–18884
     [Google Scholar]
  54. Skirrow M.B. 1982; Campylobacter enteritis - the first five years.. J Hyg 89:175–184
     [Google Scholar]
  55. Szabo M., Bronner D., Whitfield C. 1995; Relationships between rfb gene clusters required for biosynthesis of identical d-galactose-containing O antigens in Klebsiella pneumoniae serotype Ol and Serratia marcescens serotype O16.. J Bacteriol 177:1544–1553
     [Google Scholar]
  56. Taylor D.E. 1992; Genetics of Campylobacter.. Annu Rev Microbiol 46:35–64
     [Google Scholar]
  57. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets, procedure and some applications.. Proc Natl Acad Sci USA 764350–4354
     [Google Scholar]
  58. Yanisch-Perron C.J., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mpl8 and pUC19 vectors.. Gene 33:103–119
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-11-3481
Loading
Expression of Campylobacter hyoilei lipo-oligosaccharide (LOS) antigens in Escherichia coli
Microbiology 143, 3481 (1997); https://doi.org/10.1099/00221287-143-11-3481
/content/journal/micro/10.1099/00221287-143-11-3481
/content/journal/micro/10.1099/00221287-143-11-3481
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error