Abstract
Urease from ureaplasmas was purified by immunoaffinity chromatography, and the N-terminal amino acid sequence was determined for two of the three subunits. These sequences were used to design primers for a polymerase chain reaction (PCR) that amplified most of the gene coding for one of the subunits. By using a novel "PCR walking" technique, we synthesized almost the complete locus on two overlapping PCR products. We present here a partial nucleotide sequence of the urease locus from Ureaplasma urealyticum (serotype 8), which agrees with our N-terminal amino acid data but differs slightly from the sequence previously reported (A. Blanchard, Mol. Microbiol. 4:669-676, 1990). Also described are PCR primers, intended for diagnostic use, that amplify a sequence from all Ureaplasma strains tested but not from any other mycoplasmas or urease-positive bacteria.
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- Blanchard A., Barile M. F. Cloning of Ureaplasma urealyticum DNA sequences showing genetic homology with urease genes from gram-negative bacteria. Res Microbiol. 1989 May-Jun;140(4-5):281–290. doi: 10.1016/0923-2508(89)90020-x. [DOI] [PubMed] [Google Scholar]
- Blanchard A. Ureaplasma urealyticum urease genes; use of a UGA tryptophan codon. Mol Microbiol. 1990 Apr;4(4):669–676. doi: 10.1111/j.1365-2958.1990.tb00636.x. [DOI] [PubMed] [Google Scholar]
- Cassell G. H., Crouse D. T., Waites K. B., Rudd P. T., Davis J. K. Does Ureaplasma urealyticum cause respiratory disease in newborns? Pediatr Infect Dis J. 1988 Aug;7(8):535–541. [PubMed] [Google Scholar]
- Cassell G. H., Waites K. B., Crouse D. T., Rudd P. T., Canupp K. C., Stagno S., Cutter G. R. Association of Ureaplasma urealyticum infection of the lower respiratory tract with chronic lung disease and death in very-low-birth-weight infants. Lancet. 1988 Jul 30;2(8605):240–245. doi: 10.1016/s0140-6736(88)92536-6. [DOI] [PubMed] [Google Scholar]
- Christensen W. B. Urea Decomposition as a Means of Differentiating Proteus and Paracolon Cultures from Each Other and from Salmonella and Shigella Types. J Bacteriol. 1946 Oct;52(4):461–466. doi: 10.1128/jb.52.4.461-466.1946. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clayton C. L., Pallen M. J., Kleanthous H., Wren B. W., Tabaqchali S. Nucleotide sequence of two genes from Helicobacter pylori encoding for urease subunits. Nucleic Acids Res. 1990 Jan 25;18(2):362–362. doi: 10.1093/nar/18.2.362. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Delisle G. J. Multiple forms of urease in cytoplasmic fractions of Ureaplasma urealyticum. J Bacteriol. 1977 Jun;130(3):1390–1392. doi: 10.1128/jb.130.3.1390-1392.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eschenbach D. A., Buchanan T. M., Pollock H. M., Forsyth P. S., Alexander E. R., Lin J. S., Wang S. P., Wentworth B. B., MacCormack W. M., Holmes K. K. Polymicrobial etiology of acute pelvic inflammatory disease. N Engl J Med. 1975 Jul 24;293(4):166–171. doi: 10.1056/NEJM197507242930403. [DOI] [PubMed] [Google Scholar]
- Eschenbach D. A. Ureaplasma urealyticum as a cause of postpartum fever. Pediatr Infect Dis. 1986 Nov-Dec;5(6 Suppl):S258–S261. doi: 10.1097/00006454-198611010-00011. [DOI] [PubMed] [Google Scholar]
- Girgis S. I., Alevizaki M., Denny P., Ferrier G. J., Legon S. Generation of DNA probes for peptides with highly degenerate codons using mixed primer PCR. Nucleic Acids Res. 1988 Nov 11;16(21):10371–10371. doi: 10.1093/nar/16.21.10371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones B. D., Mobley H. L. Proteus mirabilis urease: nucleotide sequence determination and comparison with jack bean urease. J Bacteriol. 1989 Dec;171(12):6414–6422. doi: 10.1128/jb.171.12.6414-6422.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kalman M., Kalman E. T., Cashel M. Polymerase chain reaction (PCR) amplification with a single specific primer. Biochem Biophys Res Commun. 1990 Mar 16;167(2):504–506. doi: 10.1016/0006-291x(90)92052-2. [DOI] [PubMed] [Google Scholar]
- Kundsin R. B., Driscoll S. G., Monson R. R., Yeh C., Biano S. A., Cochran W. D. Association of Ureaplasma urealyticum in the placenta with perinatal morbidity and mortality. N Engl J Med. 1984 Apr 12;310(15):941–945. doi: 10.1056/NEJM198404123101502. [DOI] [PubMed] [Google Scholar]
- Mobley H. L., Hausinger R. P. Microbial ureases: significance, regulation, and molecular characterization. Microbiol Rev. 1989 Mar;53(1):85–108. doi: 10.1128/mr.53.1.85-108.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Muto A., Yamao F., Osawa S. The genome of Mycoplasma capricolum. Prog Nucleic Acid Res Mol Biol. 1987;34:29–58. doi: 10.1016/s0079-6603(08)60492-4. [DOI] [PubMed] [Google Scholar]
- Neagle J., De Marcucci O., Dunbar B., Lindsay J. G. Component X of mammalian pyruvate dehydrogenase complex: structural and functional relationship to the lipoate acetyltransferase (E2) component. FEBS Lett. 1989 Aug 14;253(1-2):11–15. doi: 10.1016/0014-5793(89)80919-6. [DOI] [PubMed] [Google Scholar]
- Pollack J. D., Tryon V. V., Beaman K. D. The metabolic pathways of Acholeplasma and Mycoplasma: an overview. Yale J Biol Med. 1983 Sep-Dec;56(5-6):709–716. [PMC free article] [PubMed] [Google Scholar]
- Precious B. L., Thirkell D., Russell W. C. Preliminary characterization of the urease and a 96 kDa surface-expressed polypeptide of Ureaplasma urealyticum. J Gen Microbiol. 1987 Sep;133(9):2659–2670. doi: 10.1099/00221287-133-9-2659. [DOI] [PubMed] [Google Scholar]
- Pyle L. E., Corcoran L. N., Cocks B. G., Bergemann A. D., Whitley J. C., Finch L. R. Pulsed-field electrophoresis indicates larger-than-expected sizes for mycoplasma genomes. Nucleic Acids Res. 1988 Jul 11;16(13):6015–6025. doi: 10.1093/nar/16.13.6015. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quinn P. A., Butany J., Chipman M., Taylor J., Hannah W. A prospective study of microbial infection in stillbirths and early neonatal death. Am J Obstet Gynecol. 1985 Jan 15;151(2):238–249. doi: 10.1016/0002-9378(85)90020-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Taylor-Robinson D., McCormack W. M. The genital mycoplasmas (first of two parts). N Engl J Med. 1980 May 1;302(18):1003–1010. doi: 10.1056/NEJM198005013021805. [DOI] [PubMed] [Google Scholar]
- Thirkell D., Myles A. D., Precious B. L., Frost J. S., Woodall J. C., Burdon M. G., Russell W. C. The urease of Ureaplasma urealyticum. J Gen Microbiol. 1989 Feb;135(Pt 2):315–323. doi: 10.1099/00221287-135-2-315. [DOI] [PubMed] [Google Scholar]
- Thirkell D., Myles A. D., Taylor-Robinson D. A comparison of four major antigens in five human and several animal strains of ureaplasmas. J Med Microbiol. 1990 Jul;32(3):163–168. doi: 10.1099/00222615-32-3-163. [DOI] [PubMed] [Google Scholar]
- Toth A., Lesser M. L., Brooks C., Labriola D. Subsequent pregnancies among 161 couples treated for T-mycoplasma genital-tract infection. N Engl J Med. 1983 Mar 3;308(9):505–507. doi: 10.1056/NEJM198303033080907. [DOI] [PubMed] [Google Scholar]
- Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
- Waites K. B., Rudd P. T., Crouse D. T., Canupp K. C., Nelson K. G., Ramsey C., Cassell G. H. Chronic Ureaplasma urealyticum and Mycoplasma hominis infections of central nervous system in preterm infants. Lancet. 1988 Jan 2;1(8575-6):17–21. doi: 10.1016/s0140-6736(88)91002-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Zhang H., Scholl R., Browse J., Somerville C. Double stranded DNA sequencing as a choice for DNA sequencing. Nucleic Acids Res. 1988 Feb 11;16(3):1220–1220. doi: 10.1093/nar/16.3.1220. [DOI] [PMC free article] [PubMed] [Google Scholar]