title: Cloning and expression in Escherichia coli of a synthetic DNA for hirudin, the blood coagulation inhibitor in the leech.
abstract: A 235-bp DNA coding for the leech blood coagulation inhibitor, hirudin, was chemically synthesized. The synthesis involved preparation of seven long oligodeoxyribonucleotide pairs which were assembled and cloned using a rapid and simple procedure. More than half of the transformed Escherichia coli cells expressed a biosynthetic polypeptide having biological properties which were very similar to authentic hirudin from the leech Hirudo medicinalis. To achieve efficient expression, we fused the hirudin DNA to a truncated C1 repressor gene of bacteriophage lambda to create a hybrid protein. An additional methionine at the fusion point allowed the active hirudin to be cleaved off by cyanogen bromide.
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000138:chemical synthesis; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D006629:Hirudins; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics
chemical_list: D006629:Hirudins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D004247:DNA