A large fragment approach to DNA synthesis: total synthesis of a gene for the protease inhibitor eglin c from the leech Hirudo medicinalis and its expression in E. coli
Biotechnology article published in Nucleic Acids Research (1984)
Abstract
A DNA containing the coding sequence for the proteinase inhibitor protein, eglin c, from the leech Hirudo medicinalis has been obtained by enzymatic assembly of chemically synthesized DNA fragments. The synthetic gene consists of a 232 base-pair fragment containing initiation and termination codon signals with restriction enzyme recognition sites conveniently placed for cloning into a plasmid vector. Only six oligonucleotides from 34 to 61 bases in length, sharing pairwise stretches of complementary regions at their 3'-termini, were prepared by phosphotriester solid-phase synthesis. The oligomers were annealed pairwise and converted into double stranded DNA fragments by DNA polymerase I mediated repair synthesis. The fragments were assembled by ligation, and the synthetic gene was expressed in high yield in E. coli under the transcriptional control of the E. coli tryptophan promoter. The expression product was purified to homogeneity and was shown to have similar physicochemical and identical biological properties as the authentic protein isolated from the leech.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Enzymatic assembly of chemically synthesized oligonucleotides created a synthetic gene encoding leech eglin c that was cloned and expressed at high yield in E. coli under the tryptophan promoter.
Por qué esto importa para la hirudoterapia
Este artículo informa de la síntesis química total y la expresión de un gen que codifica eglin c — el inhibidor de proteasas de Hirudo medicinalis — mediante el ensamblaje de seis oligonucleótidos sintetizados químicamente en un fragmento de 232 pares de bases con señales de codón apropiadas, su clonación bajo el promotor de triptófano de E. coli y su expresión con alto rendimiento en E. coli; el producto purificado coincidió con la proteína auténtica de la sanguijuela en sus propiedades fisicoquímicas y biológicas. Para el ámbito de ASH, esto es relevante como método de producción de un bioactivo definido del secretoma de la sanguijuela, que potencialmente permitiría un suministro escalable independiente de sanguijuelas vivas. Advertencia: se trata de un estudio de biología molecular/biotecnología/síntesis génica; no contiene datos terapéuticos, in vivo ni de hirudoterapia.
Citación
A large fragment approach to DNA synthesis: total synthesis of a gene for the protease inhibitor eglin c from the leech Hirudo medicinalis and its expression in E. coli.
Rink H, Liersch M, Sieber P, Meyer F · Nucleic acids research, 1984
Contexto clínico relacionado
Explore cómo esta investigación se conecta con la práctica clínica
Añadido a la biblioteca ASH: May 26, 2026 · Última actualización del sitio: 18 de junio de 2026