Bifunctional reporter genes: construction and expression in prokaryotic and eukaryotic cells

被引：7

作者：

Goldenkova, IV ^{[1
]}

Musiychuk, KA ^{[1
]}

Piruzian, ES ^{[1
]}

机构：

[1] Russian Acad Sci, Vavilov Inst Gen Genet, Moscow 119991, Russia

来源：

MOLECULAR BIOLOGY | 2003年 / 37卷 / 02期

基金：

俄罗斯基础研究基金会;

关键词：

reporter gene; licB; lichenase; green fluorescent protein; beta-glucuronidase; bifunctional proteins; prokaryotic and eukaryotic transformed cells;

D O I：

10.1023/A:1023310124842

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：

Bifunctional reporter proteins were constructed to combine Clostridium thermocellum lichenase (LicBM2) with Aequorea victoria green fluorescent protein (GFP) or with Escherichia coli beta-glucuronidase (GUS). The major properties of the initial proteins were preserved in the hybrid ones: LicBM2 was active at 65degreesC, GFP fluoresced, and GUS hydrolyzed its substrates. LicBM2 remained active after extension of its C or N end. Bifunctional reporter systems were shown to provide a convenient tool for studying the gene expression regulation in prokaryotic (E. coli) and eukaryotic (Saccharomyces cerevisiae, mammalian) cells, advantages of one reporter compensating for drawbacks of the other.

引用

页码：307 / 313

页数：7

共 21 条

[1] REPORTER GENES - APPLICATION TO THE STUDY OF MAMMALIAN GENE-TRANSCRIPTION [J].

ALAM, J ;

COOK, JL .

ANALYTICAL BIOCHEMISTRY, 1990, 188 (02) :245-254

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3] A BIFUNCTIONAL FUSION BETWEEN BETA-GLUCURONIDASE AND NEOMYCIN PHOSPHOTRANSFERASE - A BROAD-SPECTRUM MARKER ENZYME FOR PLANTS [J].

DATLA, RSS ;

HAMMERLINDL, JK ;

PELCHER, LE ;

CROSBY, WL ;

SELVARAJ, G .

GENE, 1991, 101 (02) :239-246

[4] A BIFUNCTIONAL ENZYME, WITH SEPARATE XYLANASE AND BETA(1,3-1,4)-GLUCANASE DOMAINS, ENCODED BY THE XYND GENE OF RUMINOCOCCUS-FLAVEFACIENS [J].

FLINT, HJ ;

MARTIN, J ;

MCPHERSON, CA ;

DANIEL, AS ;

ZHANG, JX .

JOURNAL OF BACTERIOLOGY, 1993, 175 (10) :2943-2951

[5] A thermostable Clostridium thermocellum lichenase-based reporter system for studying the gene expression regulation in prokaryotic and eukaryotic cells [J].

Goldenkova, IV ;

Musiychuk, KA ;

Piruzian, ES .

MOLECULAR BIOLOGY, 2002, 36 (05) :698-704

[6] ESTABLISHMENT OF A NORADRENERGIC CLONAL LINE OF RAT ADRENAL PHEOCHROMOCYTOMA CELLS WHICH RESPOND TO NERVE GROWTH-FACTOR [J].

GREENE, LA ;

TISCHLER, AS .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1976, 73 (07) :2424-2428

[7]

JEFFERSON RA, 1987, EMBO J, V6, P3901

[8] BETA-GLUCURONIDASE FROM ESCHERICHIA-COLI AS A GENE-FUSION MARKER [J].

JEFFERSON, RA ;

BURGESS, SM ;

HIRSH, D .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (22) :8447-8451

[9]

MUSIYCHUK KA, 2000, BIOKHIMIYA, V65, P1659

[10] Reporter gene technology: The future looks bright [J].

Naylor, LH .

BIOCHEMICAL PHARMACOLOGY, 1999, 58 (05) :749-757

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