Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Apr;62(4):1451–1456. doi: 10.1128/iai.62.4.1451-1456.1994

Molecular cloning, sequence analysis, and expression of the gene encoding the immunodominant 32-kilodalton protein of Cowdria ruminantium.

A H van Vliet 1, F Jongejan 1, M van Kleef 1, B A van der Zeijst 1
PMCID: PMC186301  PMID: 8132352

Abstract

Cowdria ruminatium, the causative agent of heartwater disease, expresses an immunodominant and conserved 32-kilodalton protein (MAP1; formerly called Cr32), which is currently in use for serodiagnosis of the disease. The gene encoding this protein, designated map1, was detected, cloned, and characterized. The gene is conserved between four different stocks of C. ruminantium originating from Senegal, Sudan, South Africa, and Zimbabwe. Homology searches revealed MAP1 to be homologous to the Anaplasma marginale surface protein MSP4, a potential protective antigen. The MAP1 protein, expressed in Escherichia coli fused with glutathione S-transferase, is specifically recognized by sera from animals infected with seven different stocks of C. ruminantium.

Full text

PDF
1451

Images in this article

1453Image
on p.1453
1454Image
on p.1454

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BOVARNICK M. R., MILLER J. C., SNYDER J. C. The influence of certain salts, amino acids, sugars, and proteins on the stability of rickettsiae. J Bacteriol. 1950 Apr;59(4):509–522. doi: 10.1128/jb.59.4.509-522.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbet A. F., Palmer G. H., Myler P. J., McGuire T. C. Characterization of an immunoprotective protein complex of Anaplasma marginale by cloning and expression of the gene coding for polypeptide Am105L. Infect Immun. 1987 Oct;55(10):2428–2435. doi: 10.1128/iai.55.10.2428-2435.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barré N., Uilenberg G., Morel P. C., Camus E. Danger of introducing heartwater onto the American mainland: potential role of indigenous and exotic Amblyomma ticks. Onderstepoort J Vet Res. 1987 Sep;54(3):405–417. [PubMed] [Google Scholar]
  4. Bezuidenhout J. D., Paterson C. L., Barnard B. J. In vitro cultivation of Cowdria ruminantium. Onderstepoort J Vet Res. 1985 Jun;52(2):113–120. [PubMed] [Google Scholar]
  5. Bleumink-Pluym N. M., van Dijk L., van Vliet A. H., van der Giessen J. W., van der Zeijst B. A. Phylogenetic position of Taylorella equigenitalis determined by analysis of amplified 16S ribosomal DNA sequences. Int J Syst Bacteriol. 1993 Jul;43(3):618–621. doi: 10.1099/00207713-43-3-618. [DOI] [PubMed] [Google Scholar]
  6. Byrom B., Yunker C. E. Improved culture conditions for Cowdria ruminantium (Rickettsiales), the agent of heartwater disease of domestic ruminants. Cytotechnology. 1990 Nov;4(3):285–290. doi: 10.1007/BF00563789. [DOI] [PubMed] [Google Scholar]
  7. Corpet F. Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res. 1988 Nov 25;16(22):10881–10890. doi: 10.1093/nar/16.22.10881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dame J. B., Mahan S. M., Yowell C. A. Phylogenetic relationship of Cowdria ruminantium, agent of heartwater, to Anaplasma marginale and other members of the order Rickettsiales determined on the basis of 16S rRNA sequence. Int J Syst Bacteriol. 1992 Apr;42(2):270–274. doi: 10.1099/00207713-42-2-270. [DOI] [PubMed] [Google Scholar]
  9. Du Plessis J. L. A method for determining the Cowdria ruminantium infection rate of Amblyomma hebraeum: effects in mice injected with tick homogenates. Onderstepoort J Vet Res. 1985 Jun;52(2):55–61. [PubMed] [Google Scholar]
  10. Du Plessis J. L., Bezuidenhout J. D., Brett M. S., Camus E., Jongejan F., Mahan S. M., Martinez D. The sero-diagnosis of heartwater: a comparison of five tests. Rev Elev Med Vet Pays Trop. 1993;46(1-2):123–129. [PubMed] [Google Scholar]
  11. Dutta S. K., Shankarappa B., Mattingly-Napier B. L. Molecular cloning and analysis of recombinant major antigens of Ehrlichia risticii. Infect Immun. 1991 Mar;59(3):1162–1169. doi: 10.1128/iai.59.3.1162-1169.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fikrig E., Barthold S. W., Marcantonio N., Deponte K., Kantor F. S., Flavell R. A. Roles of OspA, OspB, and flagellin in protective immunity to Lyme borreliosis in laboratory mice. Infect Immun. 1992 Feb;60(2):657–661. doi: 10.1128/iai.60.2.657-661.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jongejan F., Morzaria S. P., Shariff O. A., Abdalla H. M. Isolation and transmission of Cowdria ruminantium (causal agent of heartwater disease) in Blue Nile Province, Sudan. Vet Res Commun. 1984 May;8(2):141–145. doi: 10.1007/BF02214705. [DOI] [PubMed] [Google Scholar]
  14. Jongejan F. Protective immunity to heartwater (Cowdria ruminantium infection) is acquired after vaccination with in vitro-attenuated rickettsiae. Infect Immun. 1991 Feb;59(2):729–731. doi: 10.1128/iai.59.2.729-731.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jongejan F., Thielemans M. J., Brière C., Uilenberg G. Antigenic diversity of Cowdria ruminantium isolates determined by cross-immunity. Res Vet Sci. 1991 Jul;51(1):24–28. doi: 10.1016/0034-5288(91)90025-j. [DOI] [PubMed] [Google Scholar]
  16. Jongejan F., Thielemans M. J., De Groot M., van Kooten P. J., van der Zeijst B. A. Competitive enzyme-linked immunosorbent assay for heartwater using monoclonal antibodies to a Cowdria ruminantium-specific 32-kilodalton protein. Vet Microbiol. 1991 Jul;28(2):199–211. doi: 10.1016/0378-1135(91)90093-u. [DOI] [PubMed] [Google Scholar]
  17. Jongejan F., Thielemans M. J. Identification of an immunodominant antigenically conserved 32-kilodalton protein from Cowdria ruminantium. Infect Immun. 1989 Oct;57(10):3243–3246. doi: 10.1128/iai.57.10.3243-3246.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jongejan F., Uilenberg G., Franssen F. F., Gueye A., Nieuwenhuijs J. Antigenic differences between stocks of Cowdria ruminantium. Res Vet Sci. 1988 Mar;44(2):186–189. [PubMed] [Google Scholar]
  19. Jongejan F., Wassink L. A., Thielemans M. J., Perie N. M., Uilenberg G. Serotypes in Cowdria ruminantium and their relationship with Ehrlichia phagocytophila determined by immunofluorescence. Vet Microbiol. 1989 Nov;21(1):31–40. doi: 10.1016/0378-1135(89)90016-3. [DOI] [PubMed] [Google Scholar]
  20. Jongejan F., Zandbergen T. A., van de Wiel P. A., de Groot M., Uilenberg G. The tick-borne rickettsia Cowdria ruminantium has a Chlamydia-like developmental cycle. Onderstepoort J Vet Res. 1991 Dec;58(4):227–237. [PubMed] [Google Scholar]
  21. Jongejan F., de Vries N., Nieuwenhuijs J., Van Vliet A. H., Wassink L. A. The immunodominant 32-kilodalton protein of Cowdria ruminantium is conserved within the genus Ehrlichia. Rev Elev Med Vet Pays Trop. 1993;46(1-2):145–152. [PubMed] [Google Scholar]
  22. Logan L. L., Holland C. J., Mebus C. A., Ristic M. Serological relationship between Cowdria ruminantium and certain ehrlichia. Vet Rec. 1986 Nov 1;119(18):458–459. doi: 10.1136/vr.119.18.458. [DOI] [PubMed] [Google Scholar]
  23. Mahan S. M., Tebele N., Mukwedeya D., Semu S., Nyathi C. B., Wassink L. A., Kelly P. J., Peter T., Barbet A. F. An immunoblotting diagnostic assay for heartwater based on the immunodominant 32-kilodalton protein of Cowdria ruminantium detects false positives in field sera. J Clin Microbiol. 1993 Oct;31(10):2729–2737. doi: 10.1128/jcm.31.10.2729-2737.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mahan S. M., Waghela S. D., McGuire T. C., Rurangirwa F. R., Wassink L. A., Barbet A. F. A cloned DNA probe for Cowdria ruminantium hybridizes with eight heartwater strains and detects infected sheep. J Clin Microbiol. 1992 Apr;30(4):981–986. doi: 10.1128/jcm.30.4.981-986.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Oberle S. M., Barbet A. F. Derivation of the complete msp4 gene sequence of Anaplasma marginale without cloning. Gene. 1993 Dec 22;136(1-2):291–294. doi: 10.1016/0378-1119(93)90482-i. [DOI] [PubMed] [Google Scholar]
  26. Oberle S. M., Palmer G. H., Barbet A. F. Expression and immune recognition of the conserved MSP4 outer membrane protein of Anaplasma marginale. Infect Immun. 1993 Dec;61(12):5245–5251. doi: 10.1128/iai.61.12.5245-5251.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Palmer G. H., Barbet A. F., Davis W. C., McGuire T. C. Immunization with an isolate-common surface protein protects cattle against anaplasmosis. Science. 1986 Mar 14;231(4743):1299–1302. doi: 10.1126/science.3945825. [DOI] [PubMed] [Google Scholar]
  28. Palmer G. H., Oberle S. M., Barbet A. F., Goff W. L., Davis W. C., McGuire T. C. Immunization of cattle with a 36-kilodalton surface protein induces protection against homologous and heterologous Anaplasma marginale challenge. Infect Immun. 1988 Jun;56(6):1526–1531. doi: 10.1128/iai.56.6.1526-1531.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Perreau P., Morel P. C., Barre N., Durand P. Existence de la cowdriose (heartwater) à Cowdria ruminantium chez les ruminants des Antilles françaises (la Guadeloupe) et des Mascareignes (la Réunion et Ile Maurice). Rev Elev Med Vet Pays Trop. 1980;33(1):21–22. [PubMed] [Google Scholar]
  31. Rossouw M., Neitz A. W., de Waal D. T., du Plessis J. L., van Gas L., Brett S. Identification of the antigenic proteins of Cowdria ruminantium. Onderstepoort J Vet Res. 1990 Dec;57(4):215–221. [PubMed] [Google Scholar]
  32. Shankarappa B., Dutta S. K., Mattingly-Napier B. Identification of the protective 44-kilodalton recombinant antigen of Ehrlichia risticii. Infect Immun. 1992 Feb;60(2):612–617. doi: 10.1128/iai.60.2.612-617.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  34. Tebele N., McGuire T. C., Palmer G. H. Induction of protective immunity by using Anaplasma marginale initial body membranes. Infect Immun. 1991 Sep;59(9):3199–3204. doi: 10.1128/iai.59.9.3199-3204.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Van Kleef M., Neitz A. W., De Waal D. T. Isolation and characterization of antigenic proteins of Cowdria ruminantium. Rev Elev Med Vet Pays Trop. 1993;46(1-2):157–164. [PubMed] [Google Scholar]
  37. Waghela S. D., Rurangirwa F. R., Mahan S. M., Yunker C. E., Crawford T. B., Barbet A. F., Burridge M. J., McGuire T. C. A cloned DNA probe identifies Cowdria ruminantium in Amblyomma variegatum ticks. J Clin Microbiol. 1991 Nov;29(11):2571–2577. doi: 10.1128/jcm.29.11.2571-2577.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weiss E., Coolbaugh J. C., Williams J. C. Separation of viable Rickettsia typhi from yolk sac and L cell host components by renografin density gradient centrifugation. Appl Microbiol. 1975 Sep;30(3):456–463. doi: 10.1128/am.30.3.456-463.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  40. du Plessis J. L., Fourie N., Nel P. W., Evezard D. N. Concurrent babesiosis and ehrlichiosis in the dog: blood smear examination supplemented by the indirect fluorescent antibody test, using Cowdria ruminantium as antigen. Onderstepoort J Vet Res. 1990 Sep;57(3):151–155. [PubMed] [Google Scholar]
  41. du Plessis J. L., Kumm N. A. The passage of Cowdria ruminantium in mice. J S Afr Vet Med Assoc. 1971 Sep;42(3):217–221. [PubMed] [Google Scholar]
  42. van Vliet A. H., Jongejan F., van Kleef M., van der Zeijst B. A. Cloning and partial characterization of the Cr32 gene of Cowdria ruminantium. Rev Elev Med Vet Pays Trop. 1993;46(1-2):167–170. [PubMed] [Google Scholar]
  43. van Vliet A. H., Jongejan F., van der Zeijst B. A. Phylogenetic position of Cowdria ruminantium (Rickettsiales) determined by analysis of amplified 16S ribosomal DNA sequences. Int J Syst Bacteriol. 1992 Jul;42(3):494–498. doi: 10.1099/00207713-42-3-494. [DOI] [PubMed] [Google Scholar]
  44. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES