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A reporter system for prokaryotic and eukaryotic cells based on the thermostable lichenase from Clostridium thermocellum

被引：24
作者：
Piruzian, ES
Goldenkova, IV
Musiychuk, KA
Kobets, NS
Arman, IP
Bobrysheva, IV
Chekhuta, IA
Glazkova, D
机构：
[1] Russian Acad Sci, Vavilov Inst Gen Genet, Moscow 119991, Russia
[2] Russian Acad Sci, Inst Mol Genet, Moscow 123182, Russia
来源：
MOLECULAR GENETICS AND GENOMICS | 2002年 / 266卷 / 05期
基金：
俄罗斯基础研究基金会;
关键词：
reporter gene; licB; lichenase; thermostable enzyme; transcriptional and translational fusion;
D O I：
10.1007/s00438-001-0595-8
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The coding region of the licB gene from Clostridium thermocellum was truncated at the 3' end. The modified lichenase encoded by the construct (LicBM2) retained the most important properties of the enzyme - its high activity and thermostability. LicBM2 consists of the catalytic domain and part of the Pro-Thr-box. We demonstrated the application of the licBM2 gene as a reporter system for prokaryotic (Eseherichia coli) and eukaryotic (Saccharomyces cerevisiae and mammalian) cells by expressing it either as a transcriptional fusion with selected promoters or as a translational fusion with the E. coli uidA gene. The assays available for LicB activity are sensitive, accurate and simple, and can be used for the analysis of various gene fusion systems or for screening of transformants.
引用
收藏
页码：778 / 786
页数：9
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