Abstract
Repetitious gene cassettes that encode the consensus decapeptide repeat of Mytilus edulis bioadhesive protein were designed, constructed, and expressed in Escherichia coli. The bioadhesive precursor (BP) with a MW of 25,000 was expressed from one 600-bp gene comprised of a 30-bp unit repeat that accounts for E. coli codon bias. In strains employing T7 RNA polymerase for induction, BP was produced at levels approaching 60% of total cell protein. BP forms intracellular inclusions and yet methionine was processed from the N-terminus of the purified protein as shown by amino acid composition and N-terminal sequencing to give an authentic consensus precursor protein. Although the repetitious gene containing 30-bp repeat units appeared stable in T7-based host/vector systems, it was less stable in a λPL promoter-based host/vector system. Codon diversification was examined as a potential method to alleviate the problems by constructing a repetitious gene comprised of 120-bp repeats. This longer repeat unit failed to confer additional stability upon the repetitious gene.
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Salerno, A.J., Goldberg, I. (1994). Expression of a Synthetic Mussel Adhesive Protein in Escherichia Coli . In: Gebelein, C.G., Carraher, C.E. (eds) Biotechnology and Bioactive Polymers. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-9519-6_11
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DOI: https://doi.org/10.1007/978-1-4757-9519-6_11
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