Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Apr;174(7):2199–2207. doi: 10.1128/jb.174.7.2199-2207.1992

Physical and genetic mapping of the genomes of five Mycoplasma hominis strains by pulsed-field gel electrophoresis.

S A Ladefoged 1, G Christiansen 1
PMCID: PMC205839  PMID: 1551841

Abstract

We present the complete maps of five Mycoplasma hominis genomes, including a detailed restriction map and the locations of a number of genetic loci. The restriction fragments were resolved by field inversion gel electrophoresis or by the contour-clamped homogeneous-electric-field system of pulsed-field gel electrophoresis. All the ApaI, SmaI, BamHI, XhoI, and SalI restriction sites (total of 21 to 33 sites in each strain) were placed on the physical map, yielding an average resolution of 26 kb. The maps were constructed using three different approaches: (i) size determination of DNA fragments partially or completely cleaved with one or two restriction enzymes, (ii) hybridization analysis with purified restriction fragments and specific probes, and (iii) use of linking clones. A genetic map was constructed by hybridization with gene-specific probes for rpoA, rpoC, rrn, tuf, gyrB, hup, ftsY, the unc operon, the genes for two M. hominis-specific antigenic membrane proteins, and one gene encoding a protein with some homology to Escherichia coli alanyl-tRNA synthetase. The positions of mapped loci were partially conserved in the five strains except in one strain in which a 300-kb fragment was inverted. The numbers and order of mapped restriction sites were only partly conserved, and this conservation was restricted to certain regions. The gene order was compared with the gene order established for other bacteria and was found to be identical to that of the phylogenetically related Clostridium perfringens. The genome size of the M. hominis strains varied from 704 to 825 kb.

Full text

PDF
2202

Images in this article

2201Image
on p.2201

Selected References

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

  1. Andersen H., Birkelund S., Christiansen G., Freundt E. A. Electrophoretic analysis of proteins from Mycoplasma hominis strains detected by SDS-PAGE, two-dimensional gel electrophoresis and immunoblotting. J Gen Microbiol. 1987 Jan;133(1):181–191. doi: 10.1099/00221287-133-1-181. [DOI] [PubMed] [Google Scholar]
  2. Bak A. L., Black F. T., Christiansen C., Freundt E. A. Genome size of mycoplasmal DNA. Nature. 1969 Dec 20;224(5225):1209–1210. doi: 10.1038/2241209a0. [DOI] [PubMed] [Google Scholar]
  3. Bancroft I., Wolk C. P., Oren E. V. Physical and genetic maps of the genome of the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. J Bacteriol. 1989 Nov;171(11):5940–5948. doi: 10.1128/jb.171.11.5940-5948.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barile M. F., Grabowski M. W., Stephens E. B., O'Brien S. J., Simonson J. M., Izumikawa K., Chandler D. K., Taylor-Robinson D., Tully J. G. Mycoplasma hominis- tissue cell interactions: a review with new observations on phenotypic and genotypic properties. Sex Transm Dis. 1983 Oct-Dec;10(4 Suppl):345–354. [PubMed] [Google Scholar]
  5. Bautsch W. Rapid physical mapping of the Mycoplasma mobile genome by two-dimensional field inversion gel electrophoresis techniques. Nucleic Acids Res. 1988 Dec 23;16(24):11461–11467. doi: 10.1093/nar/16.24.11461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bode H. R., Morowitz H. J. Size and structure of the Mycoplasma hominis H39 chromosome. J Mol Biol. 1967 Jan 28;23(2):191–199. doi: 10.1016/s0022-2836(67)80026-3. [DOI] [PubMed] [Google Scholar]
  7. Brewer B. J. When polymerases collide: replication and the transcriptional organization of the E. coli chromosome. Cell. 1988 Jun 3;53(5):679–686. doi: 10.1016/0092-8674(88)90086-4. [DOI] [PubMed] [Google Scholar]
  8. Butler P. D., Moxon E. R. A physical map of the genome of Haemophilus influenzae type b. J Gen Microbiol. 1990 Dec;136(12):2333–2342. doi: 10.1099/00221287-136-12-2333. [DOI] [PubMed] [Google Scholar]
  9. Canard B., Cole S. T. Genome organization of the anaerobic pathogen Clostridium perfringens. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6676–6680. doi: 10.1073/pnas.86.17.6676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chang N., Taylor D. E. Use of pulsed-field agarose gel electrophoresis to size genomes of Campylobacter species and to construct a SalI map of Campylobacter jejuni UA580. J Bacteriol. 1990 Sep;172(9):5211–5217. doi: 10.1128/jb.172.9.5211-5217.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Christiansen C., Frydenberg J., Christiansen G., Andersen H., Hedegaard L. Analysis of the mycoplasma genome by recombinant DNA technology. Isr J Med Sci. 1984 Sep;20(9):781–784. [PubMed] [Google Scholar]
  12. Christiansen G., Andersen H., Birkelund S., Freundt E. A. Genomic and gene variation in Mycoplasma hominis strains. Isr J Med Sci. 1987 Jun;23(6):595–602. [PubMed] [Google Scholar]
  13. Cocks B. G., Pyle L. E., Finch L. R. A physical map of the genome of Ureaplasma urealyticum 960T with ribosomal RNA loci. Nucleic Acids Res. 1989 Aug 25;17(16):6713–6719. doi: 10.1093/nar/17.16.6713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Colman S. D., Hu P. C., Litaker W., Bott K. F. A physical map of the Mycoplasma genitalium genome. Mol Microbiol. 1990 Apr;4(4):683–687. doi: 10.1111/j.1365-2958.1990.tb00638.x. [DOI] [PubMed] [Google Scholar]
  15. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ely B., Gerardot C. J. Use of pulsed-field-gradient gel electrophoresis to construct a physical map of the Caulobacter crescentus genome. Gene. 1988 Sep 7;68(2):323–333. doi: 10.1016/0378-1119(88)90035-2. [DOI] [PubMed] [Google Scholar]
  17. Hattori M., Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. doi: 10.1016/0003-2697(86)90403-3. [DOI] [PubMed] [Google Scholar]
  18. Kauc L., Mitchell M., Goodgal S. H. Size and physical map of the chromosome of Haemophilus influenzae. J Bacteriol. 1989 May;171(5):2474–2479. doi: 10.1128/jb.171.5.2474-2479.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Krause D. C., Mawn C. B. Physical analysis and mapping of the Mycoplasma pneumoniae chromosome. J Bacteriol. 1990 Sep;172(9):4790–4797. doi: 10.1128/jb.172.9.4790-4797.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ladefoged S. A., Christiansen G. Analysis of the nucleotide sequence of the Mycoplasma hominis tuf gene and its flanking region. FEMS Microbiol Lett. 1991 Apr 15;63(2-3):133–139. doi: 10.1016/0378-1097(91)90075-l. [DOI] [PubMed] [Google Scholar]
  21. Lee J. J., Smith H. O., Redfield R. J. Organization of the Haemophilus influenzae Rd genome. J Bacteriol. 1989 Jun;171(6):3016–3024. doi: 10.1128/jb.171.6.3016-3024.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lin J. S., Kass E. H. Serological reactions of Mycoplasma hominis: differences among mycoplasmacidal, metabolic inhibition, and growth agglutination tests. Infect Immun. 1974 Sep;10(3):535–540. doi: 10.1128/iai.10.3.535-540.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McClelland M., Jones R., Patel Y., Nelson M. Restriction endonucleases for pulsed field mapping of bacterial genomes. Nucleic Acids Res. 1987 Aug 11;15(15):5985–6005. doi: 10.1093/nar/15.15.5985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Muto A. The genome structure of Mycoplasma capricolum. Isr J Med Sci. 1987 May;23(5):334–341. [PubMed] [Google Scholar]
  25. Neimark H. C. Division of mycoplasmas into subgroups. J Gen Microbiol. 1970 Oct;63(2):249–263. doi: 10.1099/00221287-63-2-249. [DOI] [PubMed] [Google Scholar]
  26. Noll K. M. Chromosome map of the thermophilic archaebacterium Thermococcus celer. J Bacteriol. 1989 Dec;171(12):6720–6725. doi: 10.1128/jb.171.12.6720-6725.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pyle L. E., Finch L. R. A physical map of the genome of Mycoplasma mycoides subspecies mycoides Y with some functional loci. Nucleic Acids Res. 1988 Jul 11;16(13):6027–6039. doi: 10.1093/nar/16.13.6027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pyle L. E., Finch L. R. Preparation and FIGE separation of infrequent restriction fragments from Mycoplasma mycoides DNA. Nucleic Acids Res. 1988 Mar 25;16(5):2263–2268. doi: 10.1093/nar/16.5.2263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pyle L. E., Taylor T., Finch L. R. Genomic maps of some strains within the Mycoplasma mycoides cluster. J Bacteriol. 1990 Dec;172(12):7265–7268. doi: 10.1128/jb.172.12.7265-7268.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rasmussen O. F., Christiansen C. Identification of the proton ATPase operon in Mycoplasma strain PG50 by heterologous hybridization. Isr J Med Sci. 1987 May;23(5):393–397. [PubMed] [Google Scholar]
  31. Ratnaningsih E., Dharmsthiti S., Krishnapillai V., Morgan A., Sinclair M., Holloway B. W. A combined physical and genetic map of Pseudomonas aeruginosa PAO. J Gen Microbiol. 1990 Dec;136(12):2351–2357. doi: 10.1099/00221287-136-12-2351. [DOI] [PubMed] [Google Scholar]
  32. Razin S. Molecular biology and genetics of mycoplasmas (Mollicutes). Microbiol Rev. 1985 Dec;49(4):419–455. doi: 10.1128/mr.49.4.419-455.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Robertson J. A., Pyle L. E., Stemke G. W., Finch L. R. Human ureaplasmas show diverse genome sizes by pulsed-field electrophoresis. Nucleic Acids Res. 1990 Mar 25;18(6):1451–1455. doi: 10.1093/nar/18.6.1451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Römling U., Grothues D., Bautsch W., Tümmler B. A physical genome map of Pseudomonas aeruginosa PAO. EMBO J. 1989 Dec 20;8(13):4081–4089. doi: 10.1002/j.1460-2075.1989.tb08592.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Smith C. L., Econome J. G., Schutt A., Klco S., Cantor C. R. A physical map of the Escherichia coli K12 genome. Science. 1987 Jun 12;236(4807):1448–1453. doi: 10.1126/science.3296194. [DOI] [PubMed] [Google Scholar]
  36. Trevino L. B., Haldenwang W. G., Baseman J. B. Expression of Mycoplasma pneumoniae antigens in Escherichia coli. Infect Immun. 1986 Jul;53(1):129–134. doi: 10.1128/iai.53.1.129-134.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ventra L., Weiss A. S. Transposon-mediated restriction mapping of the Bacillus subtilis chromosome. Gene. 1989 May 15;78(1):29–36. doi: 10.1016/0378-1119(89)90311-9. [DOI] [PubMed] [Google Scholar]
  38. Weisburg W. G., Tully J. G., Rose D. L., Petzel J. P., Oyaizu H., Yang D., Mandelco L., Sechrest J., Lawrence T. G., Van Etten J. A phylogenetic analysis of the mycoplasmas: basis for their classification. J Bacteriol. 1989 Dec;171(12):6455–6467. doi: 10.1128/jb.171.12.6455-6467.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yamao F., Muto A., Kawauchi Y., Iwami M., Iwagami S., Azumi Y., Osawa S. UGA is read as tryptophan in Mycoplasma capricolum. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2306–2309. doi: 10.1073/pnas.82.8.2306. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES