Chemical synthesis and expression of a gene coding for hirudin, the thrombin-specific inhibitor from the leech Hirudo medicinalis

• Published in: Biological chemistry Hoppe-Seyler Vol. 367; no. 8; p. 731
• Main Authors: Bergmann, C, Dodt, J, Köhler, S, Fink, E, Gassen, H G
• Format: Journal Article
• Language: English
• Published: Germany 01.08.1986
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Enzyme-Linked Immunosorbent Assay; Escherichia coli - metabolism; Gene Expression Regulation; Genes, Synthetic; Hirudins - chemical synthesis; Hirudins - metabolism; Lac Operon; Leeches - metabolism; Thrombin - antagonists & inhibitors
• ISSN: 0177-3593
• DOI: 10.1515/bchm3.1986.367.2.731

A DNA containing the coding sequence for the proteinase inhibitor protein hirudin from the leech Hirudo medicinalis has been obtained by enzymic ligation of chemically synthesized deoxyoligonucleotides. The 226 bp synthetic gene carries signals for the translation initiation and termination. Fragment synthesis was performed by the Khorana ligation method as well as by the fill-in method. Efficiencies of these two methods are compared. The synthetic gene was expressed in E. coli as a fusion protein with beta-galactosidase under the control of the lac-promoter as well as a non-hybrid protein under the control of the lambda PL-promoter. The non-hybrid expression product was shown to have similar biological properties as the authentic protein isolated from the leech.