Carcinogenesis, Teratogenesis & Mutagenesis

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Development of recombinant Lac Z gene yeast cell regulated by RNR3 promoter for screening chemical mutagens

WANG Rui-kun,GE Yi-zhi,LU Yi-xin,WU Qian-qian,LI Xiang-ming*   

  1. Preventive Medicine Department, Medical College of Yangzhou University, Yangzhou 225001, Jiangsu, China
  • Received:2013-11-25 Revised:2014-02-24 Online:2014-05-30 Published:2014-05-30


OBJECTIVE: To establish recombinant Lac Z yeast cells regulated by RNR3 in order to screen chemical mutagens. METHODS:RNR3 promoter was amplified using touch down polymerase chain reaction (PCR) from yeast genome and inserted into the yeast report vector of pMP206/ERE to construct yeast Lac Z gene report vector regulated by RNR3. This vector was transformed into the W303-1A yeast cells to construct the recombinant Lac Z gene yeast cells. These yeast cells were treated by several kinds of chemical mutagens to assess the expression of RNR3. RESULTS: Actinomycin D,mitomycin C,aphidicolin and ethidium bromide could not induce RNR3 expression. Methyl methanesulfonate,chlorambucil,cisplatin,4-nitro-N-oxidation of quinoline,bleomycin,phleomycin,5-fluorouracil,camptothecin and hydroxyurea mutagen could induce RNR3 expression in a dose-dependent manner. CONCLUSION:Recombinant yeast can be used for rapid screening of many chemical mutagens.

Key words: chemical mutagen, yeast, beta galactosidase, Lac Z gene