CAJ | 학술논문

目的：建立可筛选化学诱变物的RNR3调控重组Lac Z基因酵母细胞。方法：利用降式PCR技术从酵母基因组中扩增RNR3启动子，将其插入到pMP206/ERE酵母报告载体，构建成RNR3调控的Lac Z酵母报告载体，将该载体转入W303-1A酵母细胞，构建成RNR3调控的重组Lac Z基因酵母细胞。用数种化学诱变原作用于该重组基因酵母细胞，观察其对RNR3的诱导表达。结果：放线菌素D、丝裂霉素C、阿非迪霉素和溴乙锭不能诱导RNR3的表达，而甲磺酸甲脂、瘤可宁、顺铂、4-硝基-N-氧化喹啉、博来霉素、福莱霉素、5-氟尿嘧啶、喜树碱和羟基脲诱变原与RNR3表达有明显的剂量效应关系。结论：该重组酵母可用于对多种化学诱变原的快速筛选。
목적：건립가사선화학유변물적RNR3조공중조Lac Z기인효모세포。방법：이용강식PCR기술종효모기인조중확증RNR3계동자，장기삽입도pMP206/ERE효모보고재체，구건성RNR3조공적Lac Z효모보고재체，장해재체전입W303-1A효모세포，구건성RNR3조공적중조Lac Z기인효모세포。용수충화학유변원작용우해중조기인효모세포，관찰기대RNR3적유도표체。결과：방선균소D、사렬매소C、아비적매소화추을정불능유도RNR3적표체，이갑광산갑지、류가저、순박、4-초기-N-양화규람、박래매소、복래매소、5-불뇨밀정、희수감화간기뇨유변원여RNR3표체유명현적제량효응관계。결론：해중조효모가용우대다충화학유변원적쾌속사선。
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.