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Expressie, zuivering, kristallisatie, en enzym assays van Fumarylacetoacetate hydrolase domein-bevattende eiwitten
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
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Expressie, zuivering, kristallisatie, en enzym assays van Fumarylacetoacetate hydrolase domein-bevattende eiwitten

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23,561 Views

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10:21 min

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June 20, 2019

DOI:

10.3791/59729-v

DOI:
10.3791/59729-v

•

10:21 min
June 20, 2019

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12 Views
, , , , , , , ,
1Research Institute for Biomedical Aging Research,University of Innsbruck Austria, 2Center for Molecular Biosciences Innsbruck (CMBI),University of Innsbruck Austria, 3Division of Genetic Epidemiology,Medical University of Innsbruck Austria, 4Faculty of Chemistry, Department of Organic Chemistry,University of Vienna Austria

Summary

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Expressie en zuivering van fumarylacetoacetate hydrolase domein-bevattende eiwitten wordt beschreven met voorbeelden (expressie in E. coli, FPLC). De gezuiverde proteïnen worden gebruikt voor kristallisatie en antilichamenproductie en in dienst genomen voor enzym analyses. De geselecteerde fotometrische analyses worden voorgesteld om de multi-functionaliteit van FAHD1 als Oxaloacetaat decarboxylase en acylpyruvate hydrolase te tonen.

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Weiss, A. K. H., Holzknecht, M., Cappuccio, E., Dorigatti, I., Kreidl, K., Naschberger, A., Rupp, B., Gstach, H., Jansen-Dürr, P. Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins. J. Vis. Exp. (148), e59729, doi:10.3791/59729 (2019).

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