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Chembiochem ; 16(10): 1415-9, 2015 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-25930950

RESUMO

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.


Assuntos
Sistema Livre de Células , Colágeno/genética , Biossíntese de Proteínas , RNA Mensageiro/genética , Sistema Livre de Células/metabolismo , Colágeno/química , Escherichia coli/genética , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Luciferases/análise , Luciferases/genética , Substâncias Luminescentes/análise , Substâncias Luminescentes/metabolismo , Proteínas Ligantes de Maltose/química , Proteínas Ligantes de Maltose/genética , Multimerização Proteica , Estabilidade Proteica , RNA Mensageiro/química , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética
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