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病，由积聚的 不同诱变剂对体细胞基因的影响 流程。在许多潜在的致突变化合物中发现了 在环境中，有几种金属已被确定为致突变物 以及致癌物质。接触痕量的生理效应 自然界中普遍存在的大量金属，由于天然的 过程和环境污染，都有很好的记录。 值得注意的是，锰、钴、镉、铍、银、铬、铅、镍和铜阳离子具有 在一项旨在增加突变性的试验中被证明是 测量DNA碱基替换误差。这项研究的目的是 项目是完成对诱变剂的全面分析 十三种金属盐对DNA合成保真度的影响。这个 研究使用体外保真度测试，允许检测 在合成250期间出现的各种不同的错误 天然M13mp2中包含的核苷酸靶序列 DNA模板。到目前为止，对诱变剂的详细分析 Mn2plus和Ni2plus的效果即将完成，研究 Cr3plus和Pb2plus正在顺利进行。在请求的过程中 在资助期内，剩余的选定金属阳离子(锌+， Co2plus、Be2plus Cu2plus、Hg2plus、Sr2plus、Fe2plus和Se4plus) 将对其诱变效果进行分析。对于每个 选定金属对体外DNA的剂量效应的测量 合成和保真度，突变频率和 特定类型突变的错误率，以及构建 将进行突变谱研究。这项研究的结果 该项目将提供关于突变特异性的有价值的信息 和所选金属的剂量水平，并将用于阐明 金属诱变的机制(S)。这个项目 包括各个方面的学生。学生学习基础知识 DNA操作，斑块分析，体外DNA合成， 微生物技术、DNA序列分析、数据 危险试剂的管理和安全使用。

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.