Application of the single cell gel electrophoresis on yeast cells

被引:63
作者
Miloshev, G [1 ]
Mihaylov, M [1 ]
Anachkova, B [1 ]
机构
[1] Bulgarian Acad Sci, Inst Mol Biol, BU-1113 Sofia, Bulgaria
关键词
single cell gel electrophoresis; comet assays; Saccharomyces cerevisiae; hydrogen peroxide; methyl methanesulfonate;
D O I
10.1016/S1383-5718(01)00286-8
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
In the present paper, we have applied the single cell get electrophoresis (SCGE) assay on yeast cells treating Saccharomyces cerevisiae cells with hydrogen peroxide and methyl methanesulfonate (MMS), two DNA damaging agents. In order to overcome the problem with the yeast cell wall that prevented DNA to be extended by the electric field, we disintegrated the cell wall after embedding the cells in agarose. A characteristic picture of comets with residual nuclei and tails was observed and the length of the comet tails was dependent on the concentration of the damaging agents. Yeast cells developed comets at concentrations at least 10 times lower than the concentrations at which comets begin to appear in mammalian cells after treatment with the two genotoxic agents. The higher sensitivity of the yeast comet assay and the fact that S. cerevisiae is one of the most thoroughly studied and easy to work with eukaryotic model system suggest that the proposed method could be an useful tool for investigation of the DNA damaging activity of potential genotoxins, (C) 2002 Elsevier Science B.V All rights reserved.
引用
收藏
页码:69 / 74
页数:6
相关论文
共 16 条
[1]  
CARDENAS ME, 1990, J CELL SCI, V96, P439
[2]   THE COMET ASSAY - A COMPREHENSIVE REVIEW [J].
FAIRBAIRN, DW ;
OLIVE, PL ;
ONEILL, KL .
MUTATION RESEARCH-REVIEWS IN GENETIC TOXICOLOGY, 1995, 339 (01) :37-59
[3]   Nuclear organization and transcriptional silencing in yeast [J].
Gotta, M ;
Gasser, SM .
EXPERIENTIA, 1996, 52 (12) :1136-1147
[4]  
HALLIWELL B, 1991, FEBS LETT, V281, P259
[5]   The nature and origin of DNA single-strand breaks determined with the comet assay [J].
Horváthová, E ;
Slamenová, D ;
Hlincíková, L ;
Mandal, TK ;
Gábelová, A ;
Collins, AR .
MUTATION RESEARCH-DNA REPAIR, 1998, 409 (03) :163-171
[6]   Treatment of mammalian cells with mimosine generates DNA breaks [J].
Mikhailov, I ;
Russev, G ;
Anachkova, B .
MUTATION RESEARCH-DNA REPAIR, 2000, 459 (04) :299-306
[7]   LOW-LEVELS OF EXOGENOUS HISTONE H1 IN YEAST CAUSE CELL-DEATH [J].
MILOSHEV, G ;
VENKOV, P ;
VANHOLDE, K ;
ZLATANOVA, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (24) :11567-11570
[8]  
Moller P, 2000, CANCER EPIDEM BIOMAR, V9, P1005
[9]   DNA STRAND BREAKAGE AND DNA-STRUCTURE INFLUENCE STAINING WITH PROPIDIUM IODIDE USING THE ALKALINE COMET ASSAY [J].
OLIVE, PL ;
BANATH, JP ;
FJELL, CD .
CYTOMETRY, 1994, 16 (04) :305-312
[10]   Comet assay application in environmental monitoring: DNA damage in human leukocytes and plant cells in comparison with bacterial and yeast tests [J].
Poli, P ;
Buschini, A ;
Restivo, FM ;
Ficarelli, A ;
Cassoni, F ;
Ferrero, I ;
Rossi, C .
MUTAGENESIS, 1999, 14 (06) :547-555