BAC LIBRARY PRODUCTION FOR COMPARATIVE GENETICS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。目标是为许多物种准备高质量的BAC文库，以满足科学界对比较基因组学和功能应用的兴趣。这项工作将由两个合作团队完成，一个由劳伦斯·伯克利国家实验室(LBNL)的郑博士领导，另一个在奥克兰儿童医院研究所，由Osoegawa&Dejong博士共同指导。该应用程序包括与制备细菌人工染色体(BAC)相关的四个组成部分：1)BAC文库制备，2)克隆排列和文库复制，3)文库鉴定，以及4)提高整个过程效率和BAC文库质量的研究与开发。在第一年，该联盟将产生并鉴定12个具有10倍基因组冗余的动物BAC文库。假设对更多GBAC的需求日益增加，并提高生产效率，将分别在第2年和第3年准备并确定另外17个和22个图书馆的特征。这些文库基于当前的BAC克隆技术，将补充和扩展目前可用于比较基因组分析的BAC克隆小库。新的克隆收集将通过一套标准化的测试进行分析，包括一套经济标记的筛选、有限的BAC末端测序、BAC指纹、BAC稳定性分析和插入片段大小确定。尤其重要的是质量测试，以确保低BAC稳定性分析和插件尺寸确定。尤其重要的是质量测试，以确保低水平的克隆交叉污染。一旦图书馆通过质量控制，新的资源将以与主要应用程序一致的格式提供。为了提高整个过程的效率，将制定一套标准操作程序(SOP)，将BAC克隆从ART形式转向常规的高通量程序。预计新的BAC资源将有助于常规的高吞吐量程序。预计新的BAC资源将有助于通过比较一系列动物物种的基因来更好地了解基因的功能和进化。BAC克隆还将成为创建人类疾病动物模型的工具。广泛传播克隆人是这项提议的一个没有资金但必不可少的组成部分。图书馆信息将通过出版物和我们的主页(http://www.chori.org/bacpac).)传播

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal is to prepare BAC libraries for many species, of high quality, tailored to the scientific community's interest in comparative genomics and functional applications. The work will be done by two collaborating teams, one lead by Dr. Cheng at the Lawrence Berkeley National Laboratory (LBNL) and the other one at the Children's Hospital Oakland Research Institute, co-directed by Drs. Osoegawa & deJong. The application has four components relevant to preparing Bacterial Artificial Chromosomes (BACs): 1) BAC Library preparation, 2) Clone Arraying & Library Duplication, 3) Library Characterization, and 4) Research & Development to improve the overall process efficiency and the BAC library quality. During the first year, the consortium will generate and characterize twelve animal BAC libraries with ten-fold genome redundancy. Presuming an increasing need for additional GBACs and improved production efficiency, 17 and 22 additional libraries will be prepared and characterized in years 2 and 3 respectively. The libraries, based on current BAC cloning technology, will complement and extend the small repertoire of BAC clone collections now available for comparative genome analysis. The new clone collections will be analyzed by a standardized set of tests, including screening with a set of economic markers, limited BAC-end sequencing, BAC fingerprinting, BAC stability analysis and insert size determination. Of particular importance are quality tests to ensure low BAC stability analysis and insert size determination. Of particular importance are quality tests to ensure low levels of clonal cross contamination. Once the libraries pass the quality controls, the new resources will be made available in a format consistent with major applications. To improve the overall process efficiency, a set of Standard Operating Procedures (SOP's) will be developed to move BAC cloning away from an art form towards a routine high-throughput procedures. It is expected that the new BAC resources will contributed towards a routine-high throughput procedure. It is expected that the new BAC resources will contribute to better understanding of gene function and evolution through comparison of genes in a spectrum of animal species. The BAC clones will also become tools to create animal models for human diseases. Widespread dissemination of the clones is an unfunded but essential component of this proposal. Library information will be disseminated by publication and through our home page (http://www.chori.org/bacpac).