Expression of recombinant Hirudin in transgenic mice milk driven by the goat beta-casein promoter
Research article published in Biotechnol J (2008)
Abstract
Hirudin, isolated from the leech Hirudo medicinalis, inhibits thrombin directly and several expression systems have been used to produce recombinant Hirudin (rHirudin) for pharmaceutical purposes. A DNA fragment containing the Hirudin coding sequence and goat beta-casein secretion signal was chemically synthesized in this study. The synthetic DNA then was further constructed into a goat beta-casein expression vector for mouse transgenesis. Four lines of transgenic mice were successfully developed and one line showed a meaningful anti-thrombin activity of 40,000 anti-thrombin units (ATU)/mL in their milk. In this animal line, Hirudin mRNA was found in samples of uterus and kidney with insignificant anti-thrombin activity (</= 280 ATU/g wet tissue); however, mammary glands showed a higher activity of 780 ATU/g wet tissue. Transgenic mice showed no evident physical abnormality. The purified rHirudin was further analyzed by amino acid analysis and was found to contain a tyrosine O-sulfate residue that is absent in rHirudin expression either through Escherichia coli or yeast host systems. Experimental results demonstrated that the beta-casein-promoted Hirudin transgene could be successfully expressed in a murine model and may be applicable to large mammals such as livestock for mass production of rHirudin for pharmaceuticals.
Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.
Resumen
Transgenic mice secreted recombinant hirudin in milk at 40000 anti-thrombin units/mL; purified rHirudin contained tyrosine O-sulfate residue absent in E.
Por qué esto importa para la hirudoterapia
Este estudio produjo hirudina recombinante en la leche de ratones transgénicos utilizando una secuencia codificante de hirudina sintetizada químicamente con una señal de secreción de beta-caseína caprina, obteniendo una línea que expresaba 40.000 unidades anti-trombina (ATU)/mL en leche. El tejido de la glándula mamaria mostró 780 ATU/g de tejido húmedo, mientras que otros órganos presentaron actividad insignificante; la rHirudina purificada contenía un residuo de O-sulfato de tirosina ausente en el producto expresado en E. coli o en levadura, y los ratones transgénicos no mostraron anomalía física evidente. Para ASH, esto es relevante porque aborda la producción recombinante de hirudina, un anticoagulante clave derivado de la sanguijuela, con posible aplicabilidad a mamíferos de gran tamaño para la producción farmacéutica en masa. Advertencia: Se trata de un estudio biotecnológico de animales transgénicos como prueba de concepto; no se presentan datos de eficacia farmacológica, seguridad ni clínicos más allá de la apariencia física general.
Citación
Expression of recombinant Hirudin in transgenic mice milk driven by the goat beta-casein promoter.
Yen CH et al. · Biotechnol J, 2008
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026