EXPRESSION OF cDNA FOR MARINE ANNELID METALLOPROTEIN IN ESCHERICHIA COLI
Keywords:
cDNA, Fusion protein, Marine worm, Metalloprotein, Polychaeta, E. coliAbstract
A cDNA library of Nereis diversicolor coelomocytes was constructed in expression vector λgt 11. Selection of clones of recombined phages was performed using the immunoreactivity of an anti-metalloprotein (anti-MPII) polyclonal antibody. It is of importance to note that this selection was confirmed with the use of two anti-MPII monoclonal antibodies. Conditions for strong expression of fusion proteins by bacteria which had integrated the recombined phages, and conditions for detection of these proteins by host cells. Nevertheless, it must be mentioned that these proteins, though altered, retained their antibacterial activity.
References
- Käji J.H.R. and Nordberg M., "Metallothionein and other low molecular weight metal-binding proteins", Experientia Suppl., 34, (1979), pp. 41-124.
- Dennaï N., Dhainaut-Courtois N., Bouquegneau J.M. and Nejmeddine A., "Effets du cadmium et du mercure sur un ver marin (Nereis diversicolor O.F. Müller). Mecanismes de détoxication", C.R. Acad. Sci., Paris, 302, (1986), pp. 489-494.
- Nejmeddine A., Dhainaut-Courtois N., Baert J.L., Sautière P., Fournet B.L. and Boulenguer P., "Purification and characterization of a cadmium-binding protein from Nereis diversicolor (Annelida, Polychaeta)", Comp. Biochem. Physiol., 89C, (1988), pp. 321-326.
- Benouareth D.E., Dhainaut-Courtois N., Porchet-Henneré E. and Curgy J.J., "Traslation in a cell-free system, and in Xenopus oocytes, of mRNA which specify a cadmium-binding protein in Nereis diversicolor (Annelida, Polychaeta)", Biol. Cell., 67, (1989), pp. 167-171.
- Aviv H. and Leder P., "Purification of biologically active globulin messenger RNA by chromatography of oligothymidylic acid cellulose", Proc. Natl. Acad. Sci., USA, 69, (1972), pp. 1408-1417.
- Maniatis T., Fritsh E.F. and Sambrook J., "Molecular cloning: a laboratory manual", Cold Spring Harbor, New York, (1982), p. 545.
- Gubler U. and Hoffman B.J., "A simple and very efficient method for generating cDNA librairies", Gene, 25, (1983), pp. 263-269.
- Young R.A. and Davis R.W., "Yeast RNA polymerase II genes: isolation with antibody probes", Science, 222, (1983), pp. 778-782.
- Dhainaut-Courtois N., Nejmeddine A., Baert J.L., Dhainaut A., "Localisation immunocytochimique en microscopie électronique d’une protéine complexant le cadmium chez un ver marin (Nereis diversicolor O.F. Müller)", C.R. Acad. Sci., Paris, 305, (1987), pp. 237-241.
- Porchet-Henneré E., Nejmeddine A., Baert J.L. and Dhainaut-Courtois N., "Selective immunostaining of type 1 granulocytes of polychaeta annelid Nereis diversicolor by a monoclonal antibody against a cadmium-binding protein (MPII)", Biol. Cell., 60, (1987), pp. 259-261.
- Villareal-Ramos B., "A method for the screening of fusion protein expression by λgt11 recombinant clones without the preparation of lysogens", Nucleic Acids Res., 17, (1989), p. 6421.
- Hawkes R., Niday E. and Gorden J., "A dot-immunobinding assay for monoclonal and other antibodies", Anal. Biochem., 119, (1982), pp. 142-147.
- Towbin H., Staehelin T. and Gorden J., Electrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some application", Proc. Natl. Sci., USA, 76, (1979), pp. 4350-4354.
- Laemmli U.K., Cleavage of structural proteins during assembly of the head of bacteriophage T4", Nature, 227, (1970), pp. 680-685.
- Martson F.A.O., "The purification of eukaryotic polypeptides synthesized in Escherichia coli", Biochem. J, 240, (1986), pp. 1-12.
- Dhainaut A., Raveillon B., M’Béri M., Porchet-Henneré E. and Demuynck S., "Purification of an antibacterial protein in the coelomic fluid of Nereis diversicolor (Annelida, Polychaeta). Similitude with a cadmium-binding protein", Comp. Biochem. Physiol., 94C, (1989), pp. 555-560.
- Davis A.R., Bos T.M., Veda D.P., Nayak D. and Dowbenko R.W., "Compans: Immune response to human influenza virus hemagglutinin expressed in Escherichia coli", Gene, 21, (1983), pp. 273-284.
- Itakura K., Hirose T., Crea R. and Riggs A.D., "Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin", Science, 198, (1977), pp. 1056-1063.
- Taniguchi T., Guarente L., Roberts T.M., Kimelman D., Douhan J. and Ptashne M., Expression of the human fibroblast interferon in Escherichia coli", Proc. Natl. Sci., USA, 77, (1980), pp. 5230-5233.
- Goldschmidt R., "In vivo degradation of nonsense fragments in E. coli", Nature, 228, (1970), pp. 1151-1154.
- Jacobs F.A., Romeyer F.M., Beauchemin M. and Brousseau R., "Human metallothionein-II is synthesized as a stable membrane-localized fusion protein in Escherichia coli", Gene, 83, (1989), pp. 95-103.
- Lin Z. and Zabin I., "β-galactosidase. Rates of synthesis and degradation of incomplete chains", J. Biol. Chem., 247, (1972), pp. 2205-2211.
- Berka T., Shatzman A., Zimmerman J., Strickler J. and Rosenberg M., "Efficient expression of the yeast metallothionein gene in Escherichia coli", J. Bact., 170, (1983), pp. 21-26.
- Romeyer F.M., Jacobs F.A., Masson L., Haana Z. and Brousseau R., Bioaccumulation of heavy metals in Escherichia coli expression an inducible synthetic human metallothionein gene", J. Biotech., 8, (1988), pp. 207-220.
