EFFECTS OF SELECTED METAL SALTS ON THE FIDELITY OF DNA SYNTHESIS IN VITRO

癌症现在被认为是一种遗传性疾病， 通过各种诱变剂改变体细胞中的基因 流程. 在许多潜在的致突变化合物中， 环境中，几种金属已被确定为诱变剂， 以及致癌物质。 接触微量元素的生理效应 大量的金属，在自然界中无处不在，由于自然的 过程和环境污染，有据可查。 值得注意的是，Mn、Co、Cd、Be、Ag、Cr、Pb、Ni和Cu阳离子具有 在一项旨在 测量DNA碱基替换错误。 本研究的目的 项目是完成一个全面的分析诱变 十三种金属盐对DNA合成保真度的影响。 的 研究使用体外保真度测定，其允许检测 各种不同的错误发生在合成250 天然M13 mp2内含有的核苷酸靶序列 DNA模板 迄今为止，对致突变性的详细分析 Mn ~（2+）和Ni ~（2+）的作用已接近完成， Cr 3 plus和Pb 2 plus正在顺利进行。 在要求的 资助期内，剩余的选定金属阳离子（Zn 2 plus， Co 2+、Be 2+、Cu 2+、Hg 2+、Sr 2+、Fe 2+、Se 4+） 将分析其致突变作用。 中的每 选择的金属，体外DNA剂量效应的测量 合成和保真度，突变频率的测定， 特定类型突变的错误率，以及 将进行突变谱分析。 这项研究成果 项目将提供有关诱变特异性的有价值的信息 以及所选金属的剂量水平，并将用于阐明 金属诱发突变的机制。 这个项目 学生在各个方面。 学生学习的基础知识 DNA操作，噬斑测定，体外DNA合成， 微生物学技术，DNA序列分析，数据 管理和安全使用危险试剂。

Cancer is now regarded as a genetic disease caused by accumulated alterations of genes in somatic cells by various mutagenic processes. Of the many potential mutagenic compounds found in the environment, several metals have been identified as mutagens as well as carcinogens. The physiological effects of exposure to trace quantities of metals, ubiquitous in nature as a result of natural processes and environmental contamination, are well documented. Notably, Mn, Co, Cd, Be, Ag, Cr, Pb, Ni, and Cu cations have been shown to increase mutagenicity in an assay designed to measure DNA base substitution errors. The goal of this research project is to complete a comprehensive analysis of the mutagenic effects of thirteen metal salts on the fidelity of DNA synthesis. The research uses an in vitro fidelity assay which allows detection of a variety of different errors occurring during synthesis of a 250 nucleotide target sequence contained within a natural M13mp2 DNA template. To date, a detailed analysis of the mutagenic effects of Mn2plus and Ni2plus is nearly completed, and studies on Cr3plus, and Pb2plus are well underway. During the requested funding period, the remaining selected metal cations (Zn2plus, Co2plus, Be2plus Cu2plus Hg2plus Sr2plus, Fe2plus, and Se4plus) will be analyzed for their mutagenic effects. For each of the selected metals, measurement of dose effects on in vitro DNA synthesis and fidelity, determination of mutation frequencies and error rates for specific types of mutations, and construction of mutational spectra will be undertaken. The results of this research project will provide valuable information on mutagenic specificity and dose levels for the selected metals and will be used to elucidate the mechanism(s) of metal-induced mutagenesis. This project includes students in every aspect. Students learn fundamentals of DNA manipulations, plaque assays, in vitro DNA synthesis, microbiological techniques, DNA sequence analysis, data management, and safe use of hazardous reagents.