Summary
An oligonucleotide mixture corresponding to the codons for conserved and repeated amino acid sequences of bacterial sialidases (Roggentin et al. 1989) was used to clone a 4.3 kb PstI restriction fragment of Clostridium septicum DNA in Escherichia coli. The complete nucleotide sequence of the sialidase gene was determined from this fragment. The derived amino acid sequence corresponds to a protein of 110000 Da. The ribosomal binding site and promoter-like consensus sequences were identified upstream from the putative ATG initiation codon. The molecular and immunological properties of the sialidase expressed by E. coli are similar to those of the sialidase as isolated from C. septicum. The newly synthesized protein is assumed to include a leader peptide of 26 amino acids. On sequence alignment, the sialidases from C. septicum, C. sordellii and C. perfringens show significant homologies. As in other bacterial sialidases, conserved amino acid sequences occur at four positions in the protein. Aside from the consensus sequences, only poor homology to other bacterial and viral sialidases was found. The consensus sequence could be identified even in other, non-sialidase proteins, indicating a common function or the evolutionary relatedness of these proteins.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Berry AM, Paton JC, Glare EM, Hansmann D, Catcheside DEA (1988) Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli. Gene 71:299–305
Calza R, Huttner E, Vincentz M, Rouze P, Calangau F, Vaucheret H, Cherel I, Meyer C, Kronenberger J, Caboche M (1987) Cloning of DNA fragments complementary to tobacco nitrate reductase mRNA and encoding epitopes common to the nitrate reductases from higher plants. Mol Gen Genet 209:552–562
Cato EP, George WL, Finegold SM (1986) The clostridia. In: Sneath PHA (ed) Bergey's manual of systematic bacteriology, vol 2. Williams and Wilkins, Baltimore, pp 1141–1200
Corfield AP, Schauer R (1982) Metabolism of sialic acids. In: Schauer R (ed) Sialic acids: chemistry, metabolism and function. (Cell biology monographs, vol 10) Springer, Wien, pp 195–262
Davis LG, Dibney MD, Battey JF (1986) Basic methods in molecular biology. Elsevier, New York
Galen J, Fasano A, Ketley J, Richardson S (1990) The role of neuraminidase in Vibrio cholerae pathogenesis. Abstracts of the annual meeting of the American Society for Microbiology, Anaheim, p 41
Hahn CS, Lustig S, Strauss EG, Strauss JH (1988) Western equine encephalitis virus is a recombinant virus. Proc Natl Acad Sci USA 85:5997–6001
Harth G, Haidaris CG, So M (1987) Neuraminidase from Trypanosoma cruzi: analysis of enhanced expression of the enzyme in infectious forms. Proc Natl Acad Sci USA 84:8320–8324
Karplus PA, Walsh KA, Herriott JR (1984) Amino acid sequence of spinach ferredoxin:NADP+ oxidoreductase. Biochemistry 23:6576–6583
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132
Linn FV, Ward H, Prioli R, Pereira MEA (1990) Identification of neuraminidase in Giardia lamblia. Abstracts of the annual meeting of the American Society for Microbiology, Anaheim, p 75
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Marmur J (1961) A procedure for the isolation of DNA from microorganisms. J Mol Biol 3:208–218
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–78
Müller HE (1970) The in vitro identification and the in vivo action of Clostridium septicum neuraminidase. Zentralbl Bakteriol Parasitenkd Infektionskr Hyg AN 1 Orig 215:530–534
Oh S-J, Kim Y-C, Park Y-W, Min S-Y, Kim I-S, Kang H-S (1987) Complete nucleotide sequence of the penicillin G acylase gene and the flanking regions, and its expression in Escherichia coli. Gen 56:87–97
Pereira MEA (1983) A developmentally regulated neuraminidase activity in Tiypanosoma cruzi. Science 219:1444–1446
Roggentin P, Berg W, Schauer R (1987) Purification and characterization of sialidase from Clostridium sordellii G12. Glycoconjugate J 4:349–359
Roggentin P, Gutschker-Gdaniec G, Hobrecht R, Schauer R (1988a) Early diagnosis of clostridial gas gangrene using sialidase antibodies. Clin Chim Acta 173:251–262
Roggentin P, Rothe B, Lottspeich F, Schauer R (1988b) Cloning and sequencing of a Clostridium perfringens sialidase gene. FEBS Lett 238:31–34
Roggentin P, Rothe B, Kaper JB, Galen J, Lawrisuk L, Vimr ER, Schauer R (1989) Conserved sequences in bacterial and viral sialidases. Glycoconjugate J 6:349–353
Romeo JM, Esmon B, Zusman DR (1986) Nucleotide sequence of the myxobacterial hemagglutinin gene contains four homologous domains. Proc Natl Acad Sci USA 83:6332–6336
Rosenberg A, Schengrund C-L (1976) Sialidases. In: Rosenberg A, Schengrund C-L (eds) Plenum Press, New York, pp 295–298
Rothe B, Roggentin P, Frank R, Blöcker H, Schauer R (1989) Cloning, sequencing and expression of a sialidase gene from Clostridium sordellii G12. J Gen Microbiol 135:3087–3096
Russo TA, Thompson JS, Godoy VG, Malamy MH (1990) Cloning and expression of the Bacteriodes fragilis TAL2480 neuraminidase gene, nanH, in Escherichia coli. J Bacteriol 172:2594–2600
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Nail Acad Sci USA 74:5463–5467
Schauer R (1983) Glycosidases with special reference to the pathophysiological role of sialidases. In: Popper H, Reutter W, Gudat F, Köttgen E (eds) Structural carbohydrates of the liver. MTP Press, Lancaster, pp 83–97
Schauer R (1985) Sialic acids and their role as biological masks. Trends Biochem Sci 10:357–360
Schauer R, Sander-Wewer M, Gutschker-Gdaniec GHM, Roggentin P, Randow EA, Hobrecht R (1985) Sialidase activity in the sera of patients and rabbits with clostridial myonecrosis. Clin Chim Acta 146:119–127
Schörgendorfer K, Schwab H, Lafferty RM (1987) Nucleotide sequence of a cloned 2.5 kb PstI-EcoRI Bacillus subtilis DNA fragment coding for levanase. Nucleic Acids Res 15:9606
Southern E (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Takle GB, Young A, Snary D, Hudson L, Nicholls SC (1989) Cloning and expression of a trypomastigote-specific 85-kilodalton surface antigen gene from Trypanosoma cruzi. Mol Biochem Parasitol 37:57–64
Taussig R, Carlson M (1983) Nucleotide sequence of the yeast SUC2 gene for invertase. Nucleic Acids Res 11:1943–1954
Teufel M, Roggentin P, Schauer R (1989) Properties of sialidase isolated from Actinomyces viscosus DSM 43798. Biol Chem Hoppe-Seyler 370:435–443
Vimr ER, Lawrisuk L, Galen J, Kaper JB (1988) Cloning and expression of the Vibrio cholerae neuraminidase gene nanH in Escherichia coli. J Bacteriol 170:1495–1504
Von Heijne G (1986) A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 14:4683–4690
Watson M (1984) Compilation of published signal sequences. Nucleic Acids Res 12:5145–5164
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and bost strains: nucleotide sequences of the Ml3mp18 and pUC19 vectors. Gene 33:103–119
Young M, Minton NP, Staudenbauer WL (1989) Recent advances in the genetics of the clostridia. FEMS Microbiol Rev 63:301–326
Author information
Authors and Affiliations
Additional information
Communicated by W. Goebel
Rights and permissions
About this article
Cite this article
Rothe, B., Rothe, B., Roggentin, P. et al. The sialidase gene from Clostridium septicum: cloning, sequencing, expression in Escherichia coli and identification of conserved sequences in sialidases and other proteins. Mol Gen Genet 226, 190–197 (1991). https://doi.org/10.1007/BF00273603
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00273603