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Chembiochem ; 16(10): 1415-9, 2015 Jul 06.
Artículo en Inglés | MEDLINE | ID: mdl-25930950

RESUMEN

We have developed a collagen-mRNA platform for controllable protein production that is intended to be less prone to the problems associated with commonly used mRNA therapy as well as with collagen skin-healing procedures. A collagen mimic was constructed according to a recombinant method and was used as scaffold for translating mRNA chains into proteins. Cysteines were genetically inserted into the collagen chain at positions allowing efficient ribosome translation activity while minimizing mRNA misfolding and degradation. Enhanced green fluorescence protein (eGFP) mRNA bound to collagen was successfully translated by cell-free Escherichia coli ribosomes. This system enabled an accurate control of specific protein synthesis by monitoring expression time and level. Luciferase-mRNA was also translated on collagen scaffold by eukaryotic cell extracts. Thus we have demonstrated the feasibility of controllable protein synthesis on collagen scaffolds by ribosomal machinery.


Asunto(s)
Sistema Libre de Células , Colágeno/genética , Biosíntesis de Proteínas , ARN Mensajero/genética , Sistema Libre de Células/metabolismo , Colágeno/química , Escherichia coli/genética , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/genética , Luciferasas/análisis , Luciferasas/genética , Sustancias Luminiscentes/análisis , Sustancias Luminiscentes/metabolismo , Proteínas de Unión a Maltosa/química , Proteínas de Unión a Maltosa/genética , Multimerización de Proteína , Estabilidad Proteica , ARN Mensajero/química , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética
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