Waiting
Login-Verarbeitung ...

Trial ends in Request Full Access Tell Your Colleague About Jove
Journal / Biochemistry / Use of Recombinant Fusion Proteins in a…
JoVE Journal
Biochemistry
Author Produced

Dieser Inhalt ist Open Access.

JoVE Journal Biochemistry
Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
 

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Article DOI: 10.3791/58824-v • 19:23 min • January 16th, 2019
January 16th, 2019 •
*1,2, *1, 1, 1, 1
1Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 2Biotechnology Research Department, Gedeon Richter Plc
* These authors contributed equally

Kapitel

Summary
Automatic Translation

Here, we present the detailed procedure of a recently developed protease assay platform utilizing N-terminal hexahistidine/maltose-binding protein and fluorescent protein-fused recombinant substrates attached to the surface of nickel-nitrilotriacetic acid magnetic agarose beads. A subsequent in-gel analysis of the assay samples separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is also presented.

Tags

Recombinant Fusion Proteins Fluorescent Protease Assay Platform In-gel Renaturation High-throughput Screening Drug Discovery MTurquoise2 MApple HIV-1 Virus Protease Expression Plasmid Hexa Histadine Affinity Tag Maltose Binding Fusion Protein Tobacco Etch Virus Protease Cloning Cassette C-terminal Fluorescent Protein PacI Restriction Endonuclease NheI Restriction Endonuclease Escherichia Coli Codon Optimized Primers
Read Article

Get cutting-edge science videos from JoVE sent straight to your inbox every month.

Waiting X
Simple Hit Counter