Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Aug;59(8):2856–2858. doi: 10.1128/iai.59.8.2856-2858.1991

Streptococcus pneumoniae produces at least two distinct enzymes with neuraminidase activity: cloning and expression of a second neuraminidase gene in Escherichia coli.

M Camara 1, T J Mitchell 1, P W Andrew 1, G J Boulnois 1
PMCID: PMC258100  PMID: 1856002

Abstract

A gene from Streptococcus pneumoniae was cloned in lambda EMBL301 and then expressed in Escherichia coli, which cleaved the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-alpha-d-N-acetylneuraminic acid. The cloned gene therefore encodes an enzyme with neuraminidase activity. On the basis of restriction mapping and DNA hybridization studies, this gene could be distinguished from another pneumococcal neuraminidase gene cloned previously (A. M. Berry, J. C. Paton, E. M. Glare, D. Hansman, and D. E. A. Catcheside, Gene 71:299-305, 1988). Both neuraminidase genes were found in each of five isolates, covering at least three serotypes, of pneumococci tested.

Full text

PDF
2856

Images in this article

2857Image
on p.2857

Selected References

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

  1. Andersson B., Dahmén J., Frejd T., Leffler H., Magnusson G., Noori G., Edén C. S. Identification of an active disaccharide unit of a glycoconjugate receptor for pneumococci attaching to human pharyngeal epithelial cells. J Exp Med. 1983 Aug 1;158(2):559–570. doi: 10.1084/jem.158.2.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berry A. M., Paton J. C., Glare E. M., Hansman D., Catcheside D. E. Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli. Gene. 1988 Nov 30;71(2):299–305. doi: 10.1016/0378-1119(88)90046-7. [DOI] [PubMed] [Google Scholar]
  3. Chen J. D., Morrison D. A. Cloning of Streptococcus pneumoniae DNA fragments in Escherichia coli requires vectors protected by strong transcriptional terminators. Gene. 1987;55(2-3):179–187. doi: 10.1016/0378-1119(87)90278-2. [DOI] [PubMed] [Google Scholar]
  4. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  5. Karn J., Brenner S., Barnett L., Cesareni G. Novel bacteriophage lambda cloning vector. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5172–5176. doi: 10.1073/pnas.77.9.5172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kelly R. T., Farmer S., Greiff D. Neuraminidase activities of clinical isolates of Diplococcus pneumoniae. J Bacteriol. 1967 Jul;94(1):272–273. doi: 10.1128/jb.94.1.272-273.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kelly R., Greiff D. Toxicity of pneumococcal neuraminidase. Infect Immun. 1970 Jul;2(1):115–117. doi: 10.1128/iai.2.1.115-117.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Langridge J., Langridge P., Bergquist P. L. Extraction of nucleic acids from agarose gels. Anal Biochem. 1980 Apr;103(2):264–271. doi: 10.1016/0003-2697(80)90266-3. [DOI] [PubMed] [Google Scholar]
  9. Lathe R., Vilotte J. L., Clark A. J. Plasmid and bacteriophage vectors for excision of intact inserts. Gene. 1987;57(2-3):193–201. doi: 10.1016/0378-1119(87)90122-3. [DOI] [PubMed] [Google Scholar]
  10. Lock R. A., Paton J. C., Hansman D. Comparative efficacy of pneumococcal neuraminidase and pneumolysin as immunogens protective against Streptococcus pneumoniae. Microb Pathog. 1988 Dec;5(6):461–467. doi: 10.1016/0882-4010(88)90007-1. [DOI] [PubMed] [Google Scholar]
  11. Lock R. A., Paton J. C., Hansman D. Purification and immunological characterization of neuraminidase produced by Streptococcus pneumoniae. Microb Pathog. 1988 Jan;4(1):33–43. doi: 10.1016/0882-4010(88)90046-0. [DOI] [PubMed] [Google Scholar]
  12. O'Toole R. D., Goode L., Howe C. Neuraminidase activity in bacterial meningitis. J Clin Invest. 1971 May;50(5):979–985. doi: 10.1172/JCI106591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. O'Toole R. D., Stahl W. L. Experimental pneumococcal meningitis. Effects of neuraminidase and other pneumococcal constituents on cerebrospinal fluid in the intact dog. J Neurol Sci. 1975 Oct;26(2):167–178. doi: 10.1016/0022-510x(75)90029-5. [DOI] [PubMed] [Google Scholar]
  14. Paton J. C., Lock R. A., Hansman D. J. Effect of immunization with pneumolysin on survival time of mice challenged with Streptococcus pneumoniae. Infect Immun. 1983 May;40(2):548–552. doi: 10.1128/iai.40.2.548-552.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Scanlon K. L., Diven W. F., Glew R. H. Purification and properties of Streptococcus pneumoniae neuraminidase. Enzyme. 1989;41(3):143–150. doi: 10.1159/000469069. [DOI] [PubMed] [Google Scholar]
  16. Shoemaker N. B., Guild W. R. Destruction of low efficiency markers is a slow process occurring at a heteroduplex stage of transformation. Mol Gen Genet. 1974;128(4):283–290. doi: 10.1007/BF00268516. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Stahl W. L., O'Toole R. D. Pneumococcal neuraminidase: purification and properties. Biochim Biophys Acta. 1972 May 12;268(2):480–487. doi: 10.1016/0005-2744(72)90343-9. [DOI] [PubMed] [Google Scholar]
  19. Tanenbaum S. W., Gulbinsky J., Katz M., Sun S. C. Separation, purification and some properties of pneumococcal neuraminidase isoenzymes. Biochim Biophys Acta. 1970 Feb 11;198(2):242–254. doi: 10.1016/0005-2744(70)90057-4. [DOI] [PubMed] [Google Scholar]
  20. Tanenbaum S. W., Sun S. C. Some molecular properties of pneumoccal neuraminidase isoenzymes. Biochim Biophys Acta. 1971 Mar 23;229(3):824–828. doi: 10.1016/0005-2795(71)90301-1. [DOI] [PubMed] [Google Scholar]
  21. Walker J. A., Allen R. L., Falmagne P., Johnson M. K., Boulnois G. J. Molecular cloning, characterization, and complete nucleotide sequence of the gene for pneumolysin, the sulfhydryl-activated toxin of Streptococcus pneumoniae. Infect Immun. 1987 May;55(5):1184–1189. doi: 10.1128/iai.55.5.1184-1189.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]

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

RESOURCES