DNA CRUCIFORMS AND HUMAN DISEASE

作为宝洁公司的一名学者，比斯勒博士发现了C1抑制剂的突变 遗传性血管神经性水肿40个家系的(C1INH)基因他发现 两个似乎共享固有DNA复制的突变热点 不忠。作为他研究的一部分，他将研究突变热点 这里概述的是C1INH基因的外显子8。这一地区形成了一个 热力学稳定的阀杆环结构。发现了相似的结构 在抗凝血酶III(ATIII)人类免疫缺陷病毒TAR和 血小板驱动生长因子A基因。ATIII和C1INH也是集群 参与这些十字形结构的序列中的点突变。 他将研究这些基因形成十字形结构和 研究这些十字架对真核生物复制机制的影响。 目前，研究内在突变机制的方法是人工的。 密集的需要广泛的序列分析。比斯勒博士设计了 一种有可能选择性地只分离细菌的系统 含有发生了移码或移位的突变质粒的 诱变。这种方法将大大减少访问所需的劳动力 各种人类疾病的十字形细胞的假定致突变性 相关基因。通过组合来自突变的信息 频率和真核复制机制暂停，比斯勒博士将 更清楚地定义复制暂停在突变中的作用。更好 对涉及内在DNA不稳定性的因素的理解是 对阐明诱变的基本机制具有重要意义。这个 来自拟议研究的观察结果将直接影响到 可能用于基因治疗的稳定结构。 比斯勒博士目前的环境非常适合支持他的 持续的职业发展。他有了自己的办公室， 实验室空间。实验室里有他需要的设备。 学习，他可以使用儿童医院的核心设备 研究基金会，包括最先进的寡核苷酸合成 设施。比斯勒博士将得到这项研究的财政支持 基金会将前往休斯顿，以便向Dr。 辛登，一名顾问。比斯勒博士的毒蛇凯瑟琳·迪克森博士 在帮助年轻调查人员培养独立能力方面有丰富的经验 从事实验室调查工作，并在多个领域拥有科学专长 比斯勒博士打算研究的。比斯勒博士也将有足够的 通过参加以下活动来加深他的基础科学知识 杂志俱乐部、研讨会和课程。

As a Procter Scholar, Dr. Bissler identified mutations in the C1 inhibitor (C1INH) gene from 40 kindred with hereditary angioneurotic edema. He found two mutation hotspots which appear to share intrinsic DNA replication infidelity. The mutation hotspot he will investigate as part of his studies outlined here is in exon 8 of the C1INH gene. This region forms a thermodynamically stable stem loop structure. Similar structures are found in the Antithrombin III (ATIII) human immunodeficiency virus TAR and platelet drive growth factor A genes. Both ATIII and C1INH also cluster point mutations in the sequence which engages in these cruciform structues. He will study the ability of these genes to form cruciform structures and examine the effect of these cruciforms on eukaryotic replication machinery. Currently, the methods for studying intrinsic mutation mechanisms are labor intensive requiring extensive sequence analysis. Dr. Bissler has designed a system which has the potential to selectively isolate only bacteria harboring mutant plasmid which has undergone frameshift or point mutagenesis. This method will greatly reduce the labor required to access the putative mutagenicity of the cruciform from various human disease associated genes. By combining the information from the mutation frequencies and eukaryotic replication machinery pausing, Dr. Bissler will more clearly define the role of replication pausing in mutagenesis. Better understanding of the factors involved in intrinsic DNA instability is important to elucidate the basic mechanisms of mutagenesis. The observations from the proposed research will bear directly on the creation of stable constructs which might be used for gene therapy. Dr. Bissler's current environment is uniquely suited to support his continued career development. He has been provided with his own office and laboratory space. The laboratory contains the equipment needed for his studies, and he has access to core equipment at the Children's Hospital Research Foundation, includes state of the art oligonucleotide synthesis facilities. Dr. Bissler will be financially supported by the Research Foundation to travel to Houston in order to learn techniques from Dr. Sinden, a consultant. Dr. Bissler's snsor, Dr. Kathieen Dixon, has had extensive experience in assisting young investigators develop independent careers in laboratory investigation and has scientific expertise in areas that Dr. Bissler proposes to study. Dr. Bissler also will have ample opportunity to further his basic science knowledge by participating in journal clubs, seminars and classes.