Core--Molecular biology

这个分子生物学核心的目标是为核心成员提供分子生物学工具，以进行超级基金相关的研究。我们的成员的研究重点是研究超基金相关金属和多环芳烃（PAH）引起的毒性和致癌性，决定个体对这些有毒物质的易感性的遗传和表观遗传因素，以及参与超基金有毒物质代谢的基因和基因产物。该核心旨在建立这项研究所需的分子技术和专业知识。到目前为止，我们已经开发出的方法来确定在p53基因的序列水平上的表观遗传变化，并映射和量化金属和PAH-DNA损伤在定义的片段和单核苷酸分辨率。这两种方法涉及使用最先进的技术：连接介导的PCR（LMPCR）和DNA测序。对于绘制表观遗传的最先进的技术：连接介导的PCR（LMPCR）和执行这些复杂的实验所需的也是费力的，需要广泛的培训。在这些方法中使用的试剂（UvrABC核酸酶）也需要费力和大量的制备。此外，从这些实验中获得的结果最好由训练有素、经验丰富的研究人员解释。分子生物学核心将集中设备和专业知识，为参与该提案的所有研究人员提供高度专业化的服务，必要时可以在全国范围内提供服务。

The objective of this Molecular Biology Core is to provide Core members with molecular biology tools to pursue Superfund-related research. Our member's research is focused on studying the toxicity and carcinogenicity induced by Superfund-related metals and polycyclic aromatic hydrocarbons (PAH), the genetic and epigenetic factors in determining an individual's susceptibility to these toxicants, and genes and gene products involved in the metabolism of Superfund toxicants. This Core aims to establish molecular techniques and expertise necessary for this research. Thus far, we have developed methods to determine the epigenetic changes at the sequence level in the p53 gene, and to map and quantify metal and PAH-DNA damage at both defined fragment and single-nucleotide resolution. These two approaches involve the use of state-of-the-art technology: ligation-mediated PCR (LMPCR) and DNA sequencing. For mapping epigenetic state-of-the-art technology: ligation-mediated PCR (LMPCR) and needed to execute these sophisticated experiments are also laborious and require extensive training. The reagents (UvrABC nucleases) used in these approaches also require laborious and extensive preparation. Furthermore, the results obtained from these experiments would be best interpreted by trained, experienced researchers. The Molecular Biology Core will centralize both equipment and expertise to provide this highly specialized service to all researchers involved in this proposal, with the possibility to provide service nationwide if necessary.