Pesheva Margarita, Krastanova Olga, Staleva Liliana, Dentcheva Valentina, Hadzhitodorov Mihail, Venkov Pencho
Sofia University, Faculty of Biology, Department of Genetics, 8 Dragan Tsankov Blvd, 1421 Sofia, Bulgaria.
J Microbiol Methods. 2005 Apr;61(1):1-8. doi: 10.1016/j.mimet.2004.10.001.
An assay based on induction by carcinogens of Ty1 transposition in Saccharomyces cerevisiae is proposed. A tester strain was developed that contains a marked Ty1 element, which allows following the transposition in the genome as a whole and a mutation, which increases cellular permeability. Hypersensitivity to chemical agents, higher cell wall porosity and transformability with plasmid DNA evidenced an enhanced cellular permeability of the tester cells. The increased permeability resulted in higher sensitivity to carcinogens. The treatment with different laboratory carcinogens induced Ty1 transposition rates in the tester strain by a factor of 10 to 20, compared to the controls. The induction is not stress-generated by the cytotoxicity of carcinogens, since treatment with NaN3 at concentrations killing 50% of the cells did not increase the transposition rate. The increase of Ty1 transposition in tester cells is specific for active carcinogens and a positive response with procarcinogens was obtained only in presence of S9 mix. The Ty1 transposition test responded positively to a number of Ames-test or DEL-test negative carcinogens. The positive response of Ty1 test was statistically significant and verified in kinetics and concentration-dependent experiments. It is concluded that the Ty1 transposition test can be used, in addition to the Ames assay, as a short-term test for detection of carcinogens.
提出了一种基于致癌物诱导酿酒酵母中Ty1转座的检测方法。构建了一个测试菌株,其含有一个带有标记的Ty1元件,可用于跟踪整个基因组中的转座情况,还带有一个可增加细胞通透性的突变。对化学试剂的超敏性、更高的细胞壁孔隙率以及对质粒DNA的转化能力证明了测试细胞的细胞通透性增强。通透性增加导致对致癌物的敏感性更高。与对照相比,用不同的实验室致癌物处理可使测试菌株中的Ty1转座率提高10至20倍。这种诱导不是由致癌物的细胞毒性产生的应激反应,因为用NaN3处理使50%的细胞死亡时,转座率并未增加。测试细胞中Ty1转座的增加对活性致癌物具有特异性,只有在存在S9混合物的情况下,前致癌物才能产生阳性反应。Ty1转座试验对一些Ames试验或DEL试验阴性的致癌物有阳性反应。Ty1试验的阳性反应具有统计学意义,并在动力学和浓度依赖性实验中得到验证。结论是,除了Ames试验外,Ty1转座试验还可作为检测致癌物的短期试验。