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. 1986 Feb;51(2):699–703. doi: 10.1128/iai.51.2.699-703.1986

Molecular characterization of Chlamydia trachomatis and Chlamydia psittaci plasmids.

T Joseph, F E Nano, C F Garon, H D Caldwell
PMCID: PMC262414  PMID: 3943908

Abstract

Plasmids from Chlamydia trachomatis LGV-434 (serotype L2) and Chlamydia psittaci meningopneumonitis strain Cal-10 were cloned into the BamHI and EcoRI sites of pBR322, respectively. The recombinant plasmids pCTL2 and pCPMn, each containing an entire respective chlamydial plasmid, were transformed into Escherichia coli. The sizes of the plasmids of C. trachomatis and C. psittaci were 7.3 and 6.2 kilobases, respectively. The two plasmids were found to be distinct by restriction endonuclease analysis, DNA-DNA hybridization, and electron microscopic heteroduplex analysis. However, partial homology was observed between restriction fragments of pCTL2 and pCPMn by Southern blot analysis. Polypeptide products encoded by these plasmids were synthesized in vitro by an E. coli-directed transcription-translation system and in vivo in E. coli maxicells and minicells. None of these polypeptides was immunoreactive with anti-chlamydial sera by immunoblotting or immunoprecipitation. Based on the comparative analysis data, the C. trachomatis and C. psittaci plasmids were found to share little genetic relatedness.

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  1. Anacker R. L., Philip R. N., Williams J. C., List R. H., Mann R. E. Biochemical and immunochemical analysis of Rickettsia rickettsii strains of various degrees of virulence. Infect Immun. 1984 Jun;44(3):559–564. doi: 10.1128/iai.44.3.559-564.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Batteiger B., Newhall W. J., 5th, Jones R. B. The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. J Immunol Methods. 1982 Dec 30;55(3):297–307. doi: 10.1016/0022-1759(82)90089-8. [DOI] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Byrne G. I., Moulder J. W. Parasite-specified phagocytosis of Chlamydia psittaci and Chlamydia trachomatis by L and HeLa cells. Infect Immun. 1978 Feb;19(2):598–606. doi: 10.1128/iai.19.2.598-606.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caldwell H. D., Hitchcock P. J. Monoclonal antibody against a genus-specific antigen of Chlamydia species: location of the epitope on chlamydial lipopolysaccharide. Infect Immun. 1984 May;44(2):306–314. doi: 10.1128/iai.44.2.306-314.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caldwell H. D., Kromhout J., Schachter J. Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis. Infect Immun. 1981 Mar;31(3):1161–1176. doi: 10.1128/iai.31.3.1161-1176.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Caldwell H. D., Kuo C. C., Kenny G. E. Antigenic analysis of Chlamydiae by two-dimensional immunoelectrophoresis. I. Antigenic heterogeneity between C. trachomatis and C. psittaci. J Immunol. 1975 Oct;115(4):963–968. [PubMed] [Google Scholar]
  9. Christen A. A., Pall M. L., Manzara T., Lurquin P. F. Rapid isolation of Escherichia coli minicells by glass-fiber filtration: study of plasmid-coded polypeptides. Gene. 1983 Aug;23(2):195–198. doi: 10.1016/0378-1119(83)90051-3. [DOI] [PubMed] [Google Scholar]
  10. Clark-Curtiss J. E., Curtiss R., 3rd Analysis of recombinant DNA using Escherichia coli minicells. Methods Enzymol. 1983;101:347–362. doi: 10.1016/0076-6879(83)01026-5. [DOI] [PubMed] [Google Scholar]
  11. Collins J. Cell-free synthesis of proteins coding for mobilisation functions of ColE1 and transposition functions of Tn3. Gene. 1979 May;6(1):29–42. doi: 10.1016/0378-1119(79)90083-0. [DOI] [PubMed] [Google Scholar]
  12. Eissenberg L. G., Wyrick P. B., Davis C. H., Rumpp J. W. Chlamydia psittaci elementary body envelopes: ingestion and inhibition of phagolysosome fusion. Infect Immun. 1983 May;40(2):741–751. doi: 10.1128/iai.40.2.741-751.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grayston J. T., Wang S. New knowledge of chlamydiae and the diseases they cause. J Infect Dis. 1975 Jul;132(1):87–105. doi: 10.1093/infdis/132.1.87. [DOI] [PubMed] [Google Scholar]
  14. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  15. Hyypiä T., Larsen S. H., Ståhlberg T., Terho P. Analysis and detection of chlamydial DNA. J Gen Microbiol. 1984 Dec;130(12):3159–3164. doi: 10.1099/00221287-130-12-3159. [DOI] [PubMed] [Google Scholar]
  16. Kingsbury D. T., Weiss E. Lack of deoxyribonucleic acid homology between species of the genus Chlamydia. J Bacteriol. 1968 Oct;96(4):1421–1423. doi: 10.1128/jb.96.4.1421-1423.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Meagher R. B., Tait R. C., Betlach M., Boyer H. W. Protein expression in E. coli minicells by recombinant plasmids. Cell. 1977 Mar;10(3):521–536. doi: 10.1016/0092-8674(77)90039-3. [DOI] [PubMed] [Google Scholar]
  19. Peterson E. M., de la Maza L. M. Characterization of Chlamydia DNA by restriction endonuclease cleavage. Infect Immun. 1983 Aug;41(2):604–608. doi: 10.1128/iai.41.2.604-608.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Radloff R., Bauer W., Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. doi: 10.1073/pnas.57.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  22. Roberts T. M., Bikel I., Yocum R. R., Livingston D. M., Ptashne M. Synthesis of simian virus 40 t antigen in Escherichia coli. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5596–5600. doi: 10.1073/pnas.76.11.5596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sancar A., Wharton R. P., Seltzer S., Kacinski B. M., Clarke N. D., Rupp W. D. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. doi: 10.1016/0022-2836(81)90234-5. [DOI] [PubMed] [Google Scholar]
  24. Schachter J., Caldwell H. D. Chlamydiae. Annu Rev Microbiol. 1980;34:285–309. doi: 10.1146/annurev.mi.34.100180.001441. [DOI] [PubMed] [Google Scholar]
  25. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Zubay G. In vitro synthesis of protein in microbial systems. Annu Rev Genet. 1973;7:267–287. doi: 10.1146/annurev.ge.07.120173.001411. [DOI] [PubMed] [Google Scholar]

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