Overexpression of gtfbeta gene fragments from Streptococcus mutans encoding Glu and Cat domains in Escherichia coli

• Autorzy: Lipinski D , Zandecka-Dziubak J. , Juzwa W. , Zeyland J. , Mrozikiewicz P.M. , Slomski R.
• Języki publikacji: EN

Abstrakty

EN

Streptococcus mutans is involved in the initiation and progression of dental caries. Fragments of a gtfB gene from Streptococcus mutans encoding Glu and Cat peptides of approximately 35 and 45 kDa, respectively, were cloned from this organism and sequenced. Recombinant Glu and Cat peptides were overexpressed in Escherichia coli with C-terminal sequence containing additional six histidine residues and enterokinase recognition site. The recombinant peptides were purified from E. coli by metal affinity chromatography. These proteins will be used to prepare efficient vaccine against dental caries.

PL

Bakterie Streptococcus mutans są główną przyczyną inicjacji i rozwoju próchnicy zębów. Fragmenty genu gtß S. mutans kodujące peptydy Glu i Cat o masie cząsteczkowej 35 i 45 kDa zostały sklonowane i zsekwencjonowane. Rekombinowane peptydy Glu i Cat ulegają nadekspresji w komórkach Escherichia coli. Końce karboksylowe peptydów zostały zmodyfikowane przez dodanie sekwencji kodującej 6 reszt histydyny oraz miejsca rozpoznawanego przez enterokinazę. Rekombinowane peptydy zostały oczyszczone z bakterii E. coli metodą chromatografii powinowactwa. Oba białka zostaną wykorzystane do przygotowania skutecznej szczepionki przeciw próchnicy zębów.

Słowa kluczowe

EN

gtfbeta gene; Streptococcus mutans; transformation; Cat peptide; gene expression; glucosyltransferase; Glu peptide; cloning; immunization; Escherichia coli; caries; overexpression

• Wydawca: -
• Czasopismo: Herba Polonica
• Rocznik: 2004
• Tom: 50
• Numer: 2
• Opis fizyczny: p.78-84,fig.,ref.

Twórcy

autor

Lipinski D

• Polish Academy of Sciences, Strzeszynska 32, 60-479 Poznan, Poland

autor

Zandecka-Dziubak J.

autor

Juzwa W.

autor

Zeyland J.

autor

Mrozikiewicz P.M.

autor

Slomski R.

Bibliografia

• 1. March PD, Microbial ecology of dental plaque and its significance in health and disease. Adv Dent Res 1994; 8 (2):263-71.
• 2. Jenkinson HF, Cell surface protein receptors in oral streptococci. FEMS Microbiol Lett 1994; 121 (2): 133-40.
• 3. Haas W, Banas JA, Ligand-binding properties of the carboxyl-terminal repeat domain of Streptococcus mutans glucan-binding protein A. J Bacteriol 2000; 182 (3):728-33.
• 4. Yu H, Nakano Y, Yamashita Y, Oho T, Koga T, Effects of antibodies against cell surface protein antigen PAc-glucosyltransferase fusion proteins on glucan synthesis and cell adhesion of Streptococcus mutans. Infect Immun 1997; 65 (6):2292-8.
• 5. Smith DJ, Taubman MA, King WF, Eida S, PowellJR, EastcottJ, Immunological characteristics of a synthetic peptide associated with a catalytic domain of mutans streptococcal glucosyltransferase. Infect Immun 1994; 62 (12):5470-6.
• 6. Smith DJ, Shoushtari B, Heschel RL, King WF, Taubman MA, Immunogenicity and protective immunity induced by synthetic peptides associated with a catalytic subdomain of mutans group streptococcal glucosyltransferase. Infect Immun 1997; 65 (11):4424-30.
• 7. Mooser G, Hefta SA, Paxton RJ, Shively JE, Lee TD, Isolation and sequence of an active-site peptide containing a catalytic aspartic acid from two Streptococcus sobrinus alpha- glucosyltransferases.J Biol Chem 1991; 266 (14):8916-22.
• 8. Smith DJ, Taubman MA, Holmberg CF, EastcottJ, King WF, Ali-Salaam P, Antigenicity and immunogenicity of a synthetic peptide derived from a glucan-binding domain of mutans streptococcal glucosyltransferase. Infect Immun 1993; 61 (7):2899-905.
• 9. Mooser G, Wong C, Isolation of a glucan-binding domain of glucosyltransferase (1,6-alpha-glucan synthase) from Streptococcus sobrinus. Infect Immun 1988; 56 (4):880-4.
• 10. Taubman MA, Smith DJ, Holmberg CJ, Eastcott JW, Coimmunization with complementary glucosyltransferase peptides results in enhanced immunogenicity and protection against dental caries. Infect Immun 2000; 68 (5):2698-703.
• 11. Taubman MA, Holmberg CJ, Smith DJ, Immunization of rats with synthetic peptide constructs from the glucan-binding or catalytic region of mutans streptococcal glucosyltransferase protects against dental caries. Infect Immun 1995; 63 (8):3088-93.
• 12. Smith DJ, King WF, Barnes La, Trantolo D, Wise DL, Taubman MA, Facilitated intranasal induction of mucosal and systemic immunity to mutans streptococcal glucosyltransferase peptide vaccines. Infect Immun 2001; 69 (8):4767-73.
• 13. Jespersgaard C, Hajishengallis G, Greenway TE, Smith DJ, Russell MW, Michalek SM, Functional and immunogenic characterization of two cloned regions of Streptococcus mutans glucosyltransferase I. Infect Immun 1999; 67 (2):810-6.
• 14. Childers NK, Tong G, Mitchell S, Kirk K, Russell MW, Michalek SM, A controlled clinical study of the effect of nasal immunization with a Streptococcus mutans antigen alone or incorporated into liposomes on induction of immune responses. Infect Immun 1999; 67 (2):618-23.
• 15. SambrookJ, Fritsch EF, Maniatis T, Molecular Cloning; a Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1989.