Hostname: page-component-78c5997874-t5tsf Total loading time: 0 Render date: 2024-11-04T02:54:24.805Z Has data issue: false hasContentIssue false
  • English
  • Français

Cloning and Expression of a Synthetic Mussel Adhesive Protein in Escherichia Coli

Published online by Cambridge University Press:  15 February 2011

Anthony J. Salerno  and
Ina Goldberg
Anthony J. Salerno
Affiliation:
Allied-Signal Inc., 101 Columbia Road, Morristown, NJ 07962.
Ina Goldberg
Affiliation:
Allied-Signal Inc., 101 Columbia Road, Morristown, NJ 07962.
Get access

Abstract

Repetitious gene cassettes that encode the consensus decapeptide repeat of Mytilus edulis bioadhesive protein were cloned and expressed in Escherichia coli. The bioadhesive precursor (BP, Mx−25,000) was expressed from one 600-bp gene comprised of a 30-bp unit repeat. The repetitious gene appeared stable in a T7-based host/vector system.

Using the T7 expression system for induction, BP was produced at levels approaching 60% of total cell protein. BP was found both in intracellular inclusions and in the soluble fraction. Interestingly, methionine was processed from the N-terminus of the purified protein to give an authentic consensus precursor protein.

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 292: Symposium S – Biomolecular Materials , 1992 , 99
Copyright
Copyright © Materials Research Society 1993

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1. Waite, J. H., Biol. Rev. 58, 209 (1983).Google Scholar
2. Waite, J. H., Comp. Biochem. Physiol. 97B, 19 (1990).Google Scholar
3. Waite, J. H., J. Biol. Chem. 258, 2911 (1983).Google Scholar
4. Waite, J. H., CHEMTECH 17, 692 (1987).Google Scholar
5. Filpula, D. R., Lee, S. M., Link, R. P., Strausberg, S. L., and Strausberg, R. L., Biotechnol. Prog. 6, 171 (1990).Google Scholar
6. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. J., Smith, J. A., and Struhl, K., Current Protocols in Molecular Biology (Greene Publishing Associates, New York, 1987).Google Scholar
7. Williams, Jon I., Salerno, A. J., Goldberg, I., and McAllister, W. T., U. S. Patent No. 5089406 (18 February 1992).Google Scholar
8. Studier, F. W., Rosenberg, A. H., Dunn, J. J., and Dubendorff, J. W., Meth. Enzymol. 185, 60 (1990).Google Scholar
9. Panyim, S. and Chalkey, R., Arch. Biochem. Biophys. 130, 337 (1969).Google Scholar
10. Meltzer, N. M., Tous, G. I., Gruber, S., and Stein, S., Anal. Biochem. 160, 356 (1987).Google Scholar
11. Debenham, P. G., TIBTECH, 10, 96 (1992).Google Scholar
12. Goldberg, I. and Salerno, A. J. in Materials Synthesis Utilizing Biological Processes, edited by Rieke, P. C., Calvert, P. D., and Alper, M. (Mater. Res. Soc. Proc. 174, Pittsburgh, PA, 1990) pp. 229–236.Google Scholar