A New Reporter Gene System Suited for Cell-Free Protein Synthesis and High-Throughput Screening in Small Reaction Volumes
✍ Scribed by René Hempel; Frank Wirsching; Andreas Schober; Andreas Schwienhorst
- Publisher
- Elsevier Science
- Year
- 2001
- Tongue
- English
- Weight
- 147 KB
- Volume
- 297
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
The properties of M-hirudin as a new reporter gene system were examined using rabbit reticulocyte lysate for cell-free protein expression. In contrast to the luciferase gene, in vitro translation of M-hirudin is highly robust against changes in concentrations of K ؉ (and Rb ؉ ). In addition, M-hirudin can be detected very sensitively using a reasonably priced fluorimetric thrombin assay. To show that the new reporter gene system is well suited for (u)HTS-applications, cell-free synthesis as well as the fluorimetric assay of M-hirudin were carried out in nanotiter and microtiter plates, respectively.