A new reporter gene system suited for cell-free protein synthesis and high-throughput screening in small reaction volumes

被引：7
|
作者：
Hempel, R
Wirsching, F
Schober, A
Schwienhorst, A
机构：
[1] Inst Mikrobiol & Genet, Abt Mol Genet & Praeparat Mol Biol, D-37077 Gottingen, Germany
[2] Inst Phys Hochtechnol, D-07743 Jena, Germany
来源：
ANALYTICAL BIOCHEMISTRY | 2001年 / 297卷 / 02期
关键词：
in vitro translation; hirudin; luciferase reporter gene; cation concentration;
D O I：
10.1006/abio.2001.5322
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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. (C) 2001 Academic Press.
引用
收藏
页码：177 / 182
页数：6
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