SEQUENCING MOUSE GENOMIC CLONES

描述（改编自研究者摘要）： 该项目的研究是系统地对 与人类基因组测序区域同线的小鼠基因组。 测序将通过延续罗伊博士的 实验室，位于俄克拉荷马州大学化学系， 生物化学在诺曼校园，在与实验室的合作， 博士约翰霍普金斯大学的里夫斯。 在过去的一年里，这种非正式的 合作已经产生了超过200 kbp的小鼠 与人类22 q11同线的染色体DNA。 罗伊医生现在希望 使这种合作正式化，并开始从他的 通过共同协调努力，开始小鼠基因组 更大规模的测序，并将此序列数据提供给 社区 全长基因组克隆测序将在 和俄克拉荷马州大学的基因组克隆文库和末端测序 将在约翰霍普金斯大学演出。 为了实现这一目标， 调查人员现在建议： 1. 扩大密歇根大学的兆碱基测序能力 俄克拉荷马州基因组技术高级中心将包括一个专门的小组 to mouse小鼠genomic基因测序. ACGT是建立在毕业生 基于学生/博士后培训的基因组测序方法， 成功地完成了大约200万个人类DNA碱基 序列和大约1倍的化脓性链球菌覆盖率 和淋病奈瑟菌的基因组 通过使用和 完善他们已经建立的协议，调查人员可以完成 01年和02年至少6个含有BAC小鼠基因组的序列 利用已经可用的NSF EPSCoR资助的ABI 377， 03年和04年的测序率， 测序仪 因此，他们将测序大约4.5 M的小鼠基因组， 与同时测序的人类基因组区域同线 4年的融资期限。 2. 继续发展、改进和实施自动化程序 用于DNA分离、DNA序列分析、数据采集和数据分析 分析，从而提高其DNA测序效率。 3. 快速向社区发布注释的DNA序列数据 继C. elegans范式 利用现有技术，调查人员可以清楚地记录 这个项目的可行性，根据自己的生产力， 华盛顿大学（C. elegans）和桑格中心（酵母，C. elegans和human）。 目前，最后序列的每个基地的成本为 略低于1美元，这是通过将NCHGR的总资金用于 调查员费用，包括间接费用和按比例分摊的设备费用， 总授予期，除以存放在 与GenBank 随着技术和效率的适度提高， 研究人员可以实际地预期， 基因组DNA可以在4年的拟议资助期内测序， 成本低于50美分每基地的最终序列。 在此期间， 他们还将继续培养下一代科学家， 发展新的测序方法所需的基本理论和方法， 到数据分析。

DESCRIPTION (Adapted from the Investigator's Abstract): The major goal of the research in this project is to systematically sequence regions of the mouse genome that are syntenic with sequenced regions of the human genome. The sequencing will be accomplished by continuing the link between Dr. Roe's laboratory, based at the University of Oklahoma Department of Chemistry and Biochemistry on the Norman campus, in a partnership with the laboratory of Dr. Reeves at John Hopkins University. During the past year, this informal collaboration has resulted in the sequence of over 200 kbp of mouse chromosomal DNA that is syntenic with human 22q11. Dr. Roe now wishes to formalize this collaboration and begin transferring the technology from his group to John Hopkins by jointly coordinating efforts to begin mouse genomic sequencing on a larger scale and provide this sequence data to the community. Full length genomic clone sequencing will be performed at the University of Oklahoma and the genomic clone libraries and end sequencing will be performed at the John Hopkins University. To achieve this goal, the investigators now propose to: 1. Expand the megabase sequencing capabilities at the University of Oklahoma Advanced Center for Genome Technology to include a group dedicated to mouse genomic sequencing. ACGT is built upon the graduate student/postdoctoral training-based approach to genomic sequencing that has led to successfully completing approximately 2 million bases of human DNA sequence and approximately one-fold coverage of the Streptococcus pyogenes and Neisseria gonorrhoeae genomes within the last year. By employing and refining their already established protocols, the investigators can complete the sequence of at least 6 mouse genomic containing BACs in years 01 and 02 with an already available NSF EPSCoR funded ABI377 and double their sequencing rate in years 03 and 04 with an additional automated DNA sequencer. Thus, they will sequence approximately 4.5 M of the mouse genome that is syntenic with concurrently sequenced human genomic regions within the proposed 4 year funding period. 2. To continue to develop, improve and implement the automated procedures for DNA isolation, DNA sequence analysis, data acquisition, and data analysis, thereby increasing their DNA sequencing efficiency. 3. To rapidly release annotated DNA sequence data to the community following the C. elegans paradigm. With the existing technology, the investigators can clearly document the feasibility of this project, based on their own productivity, that of the Washington University (C. elegans), and the Sanger Center (yeast, C. elegans, and human). The present cost per base of final sequence is slightly under $1 as calculated by taking the total NCHGR funding to the investigator, including indirect cost and pro-rating equipment over the total grant period, and dividing by the total number of bases deposited in GenBank. With the modest improvement in technology and efficiency, the investigators can realistically expect that a significant region of mouse genomic DNA can be sequenced within the 4 year proposed grant period at a cost of under 50 cents per base of final sequence. Throughout this period, they also will continue to train the next generation of scientists in the basic theories and methods needed to evolve new approaches to sequencing and to data analysis.