title: Cloning and expression of a porcine prorelaxin gene in E. coli.
abstract: A porcine prorelaxin gene has been constructed partly by synthetic means and partly from its natural messenger RNA. A gene coding for the 32 N-terminal amino acids including a chain initiator methionine codon (B gene) was synthesised and inserted in a plasmid at a site downstream from a tryptophan promoter in such a way that its expression is under the control of the trp promoter. DNA corresponding to the rest of the prorelaxin was prepared using reverse transcriptase extension of a primer complementary to relaxin mRNA and joined at a suitable restriction site to the B gene. Transformation of E. coli with this plasmid followed by suitable induction resulted in the synthesis of a new protein identified as prorelaxin by its size and its antigenic similarity to relaxin.
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D004247:DNA / Q000378:metabolism; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D046911:Macromolecular Substances; D009876:Operon; D010957:Plasmids; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D012194:RNA-Directed DNA Polymerase; D012065:Relaxin / Q000235:genetics*; D013552:Swine; D014158:Transcription, Genetic
chemical_list: D003062:Codon; D046911:Macromolecular Substances; D011498:Protein Precursors; D012333:RNA, Messenger; C038656:prorelaxin; D012065:Relaxin; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D004262:DNA Restriction Enzymes