Reporter enzyme

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A reporter enzyme is an enzyme that is used to amplify signals in a biochemical or genetic detection . This can e.g. B. be an immunoconjugate , part of a fusion protein or the gene expression product of a reporter gene .

properties

The reporter enzymes are the most commonly used dye-forming enzymes alkaline phosphatase (engl. Alkaline phosphatase , AP), the horseradish peroxidase (engl. Horseradish peroxidase , HRP), the glucose oxidase (GOX). Several reporter systems are used in the course of double immunolabeling .

As a reporter gene-based enzymes that are chemiluminescence -releasing luciferase , the chloramphenicol acetyltransferase (engl. Chloramphenicol acetyl transferase , CAT), the β-galactosidase (engl. Β-galactosidase , β-gal, the gene LacZ , known from the blue White selection ) and β-glucuronidase are used. A seldom used peculiarity is the indigoidin synthesis by a combination of the Streptomyces lavendulae non-ribosomal peptide synthase BpsA and the Streptomyces verticillus 4'-phosphopantetheinyl transferase Svp , because it works without the addition of substrate.

Fluorescent proteins such as B. GFP , roGFP , rxYFP , HyPer , VSFP and Synapto-pHluorin are reporter proteins, but mostly not enzymes.

history

Reporter enzyme-coupled antibodies were published simultaneously in 1966 by PK Nakane and GB Pierce as well as S. Avrameas and J. Uriel.

literature

  • Friedrich Lottspeich , Haralabos Zorbas: Bioanalytics . Spektrum Akademischer Verlag, Heidelberg 1998, ISBN 978-3827400413 .
  • Hubert Rehm , Thomas Letzel: The Experimenter: Protein Biochemistry / Proteomics . 6th edition, Spektrum Akademischer Verlag, Heidelberg 2009, ISBN 978-3827423122 .
  • Cornel Mülhardt: The Experimenter: Molecular Biology / Genomics. Sixth edition. Spektrum Akademischer Verlag, Heidelberg 2008, ISBN 3-82-742036-9 .

Individual evidence

  1. ↑ Steven R. Kain, Subinay Ganguly: Overview of Genetic Reporter Systems. . In: Current Protocols in Molecular Biology . 9, 2001. doi : 10.1002 / 0471142727.mb0906s36 .
  2. ↑ M. Müller, S. Ausländer, D. Ausländer, C. Kemmer, M. Fussenegger: A novel reporter system for bacterial and mammalian cells based on the non-ribosomal peptide indigoidine. In: Metabolic engineering. Volume 14, Number 4, July 2012, pp. 325-335, doi : 10.1016 / j.ymben.2012.04.002 . PMID 22543310 .
  3. ↑ PK Nakane, GB Pierce: Enzyme-labeled antibodies: preparation and application for the localization of antigens. In: The journal of histochemistry and cytochemistry: official journal of the Histochemistry Society. Volume 14, Number 12, December 1966, pp. 929-931, doi : 10.1177 / 14.12.929 , PMID 17121392 .
  4. ↑ S. Avrameas, J. Uriel, Method de marquage d'antigenes et d'anticorps avec des enzymes et son application en immune-diffusion. CR Acad. Sci. (Paris) 262: 2543 (1966).