Cloning, expression, and characterization of a synthetic analog to the bioadhesive precursor protein of the sea mussel Mytilus edulis.
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 relative molecular mass of 25,000 was expressed from one 600-bp gene at levels approaching 60% of total cell protein in strains employing T7 RNA polymerase for induction and carrying a repetitious gene comprised of a 30-bp unit repeat that accounts for E. coli codon bias. 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.
Salerno
A J
AJ
Biotechnology Department, Allied Signal Inc., Morristown, NJ 07962.
Goldberg
I
I
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D049872:Bivalvia / Q000235:genetics*; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004926:Escherichia coli; D015964:Gene Expression Regulation, Bacterial; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics* / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid
D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C082021:bioadhesive precursor protein, Mytilus