DNA CRUCIFORMS AND HUMAN DISEASE

作为Procter学者，Bissler博士鉴定了C1抑制剂中的突变， （C1INH）基因的检测。他发现 两个突变热点似乎共享内在DNA复制， 不忠作为研究的一部分， 在C1INH基因的外显子8中。该地区形成了一个 茎环结构稳定。发现了类似的结构 抗凝血酶III（ATIII）人类免疫缺陷病毒TAR， 血小板驱动生长因子A基因。ATIII和C1INH也聚集在一起 在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的能力， 研究这些十字形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劳动 需要大量的序列分析。比斯勒博士设计了 一种有可能只选择性地分离细菌的系统， 携带发生移码或点突变的突变质粒 诱变这种方法将大大减少访问 来自各种人类疾病的十字形的推定致突变性 相关基因通过结合突变的信息 频率和真核生物复制机制暂停，比斯勒博士将 更清楚地定义了复制暂停在诱变中的作用。更好 对内在DNA不稳定性相关因素的理解， 重要的是阐明诱变的基本机制。的 从拟议的研究中观察到的结果将直接影响到 稳定的构建体，可能用于基因治疗。 博士Bissler目前的环境非常适合支持他 持续的职业发展。他有自己的办公室， 实验室空间实验室里有他所需要的设备。 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Bissler博士将得到研究的财政支持 基金会前往休斯顿，以学习技术博士。 Sinden，顾问。比斯勒医生的父亲凯瑟琳狄克逊医生 在帮助年轻研究人员发展独立研究方面拥有丰富的经验 从事实验室调查，并在以下领域拥有科学专长： 比斯勒博士打算研究的东西比斯勒博士也将有足够的 有机会通过参与进一步他的基础科学知识， 期刊俱乐部、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.