title: Cloning and expression of a collagen-analog-encoding synthetic gene in Escherichia coli.
abstract: A family of totally synthetic genes coding for multiple tandem repeats of the amino acid sequence (Gly-Pro-Pro) has been prepared and inserted into the ClaI cloning site of the expression vector pJL6. A representative recombinant plasmid, pAC1, with an insert of about 340 bp was established in an Escherichia coli strain bearing a defective lambda prophage, to study expression of the CII-collagen analog fusion protein produced from pAC1 upon heat induction. Authentic fusion protein production was demonstrated by nucleotide sequencing, Northern-blot analysis, and in vivo synthesis. Conversion of a wild-type rpoH allele to the rpoH165 mutation was shown to suppress proteolysis of the unstable fusion protein.
authors.affiliation: Allied-Signal, Inc., Biosciences Program, Morristown, NJ 07960.
authors.forename: I
authors.initials: I
authors.lastname: Goldberg
authors.forename: A J
authors.initials: AJ
authors.lastname: Salerno
authors.forename: T
authors.initials: T
authors.lastname: Patterson
authors.forename: J I
authors.initials: JI
authors.lastname: Williams
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics*; D011108:Polymers; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D014158:Transcription, Genetic
chemical_list: D010455:Peptides; D011108:Polymers; D012333:RNA, Messenger; C061813:glycine-proline-proline polymer; D003094:Collagen