转染野生型DNA聚合酶β对交链孢霉酚致突变作用的影响  

Effects of wild DNA polβ transfection on the mutagenesis induced by alternariol

作　　者：张文靖[1] 赵国强[2] 赵勤[3] 杨洪艳[1] 牛静[4] 董子明[1] 

机构地区：[1]郑州大学基础医学院病理生理学教研室,郑州450001 [2]郑州大学基础医学院微生物学与免疫学教研室,郑州450001 [3]郑州大学基础医学院生物化学教研室,郑州450001 [4]河南中医学院基础医学院,郑州450008

出　　处：《郑州大学学报（医学版）》2008年第1期50-52,共3页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目39870287;教育部科学技术研究重点项目02088

摘　　要：目的:观察转染野生型DNA聚合酶β(polymerase beta,polβ)对交链孢霉酚(alternariol,AOH)致突变作用的影响。方法:用pEGFP-C3(空载体)、pEGFP-C3-polβ(野生型polβ重组表达载体)质粒转染NIH3T3细胞;转染后的细胞分别用1.5μmol/L、2.0μmol/L和2.5μmol/L3种浓度的AOH诱导48h,RT-PCR扩增DNA polβ基因,克隆至pEGM-T载体后,转化大肠杆菌JM109,筛选鉴定,进行测序分析。结果:转染pEGFP-C3的NIH3T3细胞DNA polβ基因突变点个数随AOH浓度升高而增多,而转染pEGFP-C3-polβ的NIH3T3细胞DNA polβ基因在1.5μmol/L和2.0μmol/L浓度诱导下未见突变发生,当AOH浓度增大到2.5μmol/L才出现DNA polβ基因突变。结论:高表达野生型DNA polβ能够在一定程度上抵抗霉菌毒素的致突变作用。Aim： To observe the effects of wild DNA polβ transfection on the mutagenesis induced by ahernariol （AOH）.Methods： pEGFP-C3 and pEGFP-C3-polβ were transfected respectively into NIH3T3 cell lines with lipofectamine method, and screened with G418. Two stable NIH3T3 cell lines were obtained carrying pEGFP-C3-polβ and pEGFP-C3, respectively. Two groups of NIH3T3 cells （with pEGFP-C3 and with pEGFP-C3-polβ） were induced by 1.5 μmol/L, 2.0μmol/L, and 2.5 μmol/L AOH for 48 hours,respectively. Total RNA of the cells was extracted, and then a eDNA fragment was obtained through RT-PCR technique. The PCR product was cloned into pEGM-T Easy, then sequenced. Results： The number of mutations increased following the increase of AOH concentration in the group of NIH3T3 cells carrying pEGFP-C3, while in the group of NIH3T3 cells carrying pEGFP-C3-pol[3, there was no mutation when induced by 1.5 μmol/L and 2.0 μmol/L AOH,and there were mutations when the concentration of AOH was as high as 2.5 μmol/L. Conclusion：Wild DNA polβ can inhibit mutagenesis induced by. AOH to some extent.

关 键 词：DNA聚合酶Β 霉菌毒素 突变 序列分析 

分 类 号：R730[医药卫生—肿瘤]