Realization of β-lactamase as a versatile fluorogenic reporter

被引:33
作者
Campbell, RE [1 ]
机构
[1] Univ Alberta, Dept Chem, Edmonton, AB T6G 2G2, Canada
关键词
D O I
10.1016/j.tibtech.2004.03.012
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
beta-Lactamase has emerged as the heir apparent to beta-galactosidase as a catalytic reporter for imaging biological events in live mammalian cells. In recent years, several publications have demonstrated the advantages of beta-lactamase as a reporter in applications ranging from monitoring of gene transcription to detection of protein-protein interactions. Now, Rao et al. have demonstrated that beta-lactamase can also serve as a sensitive fluorogenic reporter for imaging Tetrahymena ribozyme activity in live cells. This assay should pave the way for the screening of large libraries of ribozyme mutants by flow cytometry and, therefore, the isolation of variants with improved splicing activity in the cytoplasm of mammalian cells.
引用
收藏
页码:208 / 211
页数:4
相关论文
共 29 条
[1]   High content screening applied to large-scale cell biology [J].
Abraham, VC ;
Taylor, DL ;
Haskins, JR .
TRENDS IN BIOTECHNOLOGY, 2004, 22 (01) :15-22
[2]   Extended-spectrum β-lactamases in the 21st century:: Characterization, epidemiology, and detection of this important resistance threat [J].
Bradford, PA .
CLINICAL MICROBIOLOGY REVIEWS, 2001, 14 (04) :933-951
[3]   A monomeric red fluorescent protein [J].
Campbell, RE ;
Tour, O ;
Palmer, AE ;
Steinbach, PA ;
Baird, GS ;
Zacharias, DA ;
Tsien, RY .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (12) :7877-7882
[4]   A sensitive and specific enzyme-based assay detecting HIV-1 virion fusion in primary T lymphocytes [J].
Cavrois, M ;
de Noronha, C ;
Greene, WC .
NATURE BIOTECHNOLOGY, 2002, 20 (11) :1151-1154
[5]   FACS-optimized mutants of the green fluorescent protein (GFP) [J].
Cormack, BP ;
Valdivia, RH ;
Falkow, S .
GENE, 1996, 173 (01) :33-38
[6]   Flow cytometric screening of cell-based libraries [J].
Daugherty, PS ;
Iverson, BL ;
Georgiou, G .
JOURNAL OF IMMUNOLOGICAL METHODS, 2000, 243 (1-2) :211-227
[7]   β-Lactamase protein fragment complementation assays as in vivo and in vitro sensors of protein-protein interactions [J].
Galarneau, A ;
Primeau, M ;
Trudeau, LE ;
Michnick, SW .
NATURE BIOTECHNOLOGY, 2002, 20 (06) :619-622
[8]   Novel fluorogenic substrates for imaging 6-lactamase gene expression [J].
Gao, WZ ;
Xing, BG ;
Tsien, RY ;
Rao, JH .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (37) :11146-11147
[9]   Fluorescence readouts in HTS: no gain without pain? [J].
Gribbon, P ;
Sewing, A .
DRUG DISCOVERY TODAY, 2003, 8 (22) :1035-1043
[10]   Imaging Tetrahymena ribozyme splicing activity in single live mammalian cells [J].
Hasegawa, S ;
Jackson, WC ;
Tsien, RY ;
Rao, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (25) :14892-14896