GENOME PHYSICAL MAP OF DROSOPHILA

果蝇基因组物理图谱

• 批准号: 6564773
• 负责人: MICHAEL J PALAZZOLO
• 金额: $ 148.12万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564773
• 关键词: Drosophilidae; artificial chromosomes; computer assisted sequence analysis; gel electrophoresis; genetic library; genetic mapping; genetic markers; genome; in situ hybridization; molecular cloning; nucleic acid sequence; polymerase chain reaction; restriction fragment length polymorphism; sequence tagged sites

The initial Drosophila Genome Center grant proposed to build a clone-based physical map of the Drosophila melanogaster euchromatic genome using a bacteriophage P1 library and a non-random mapping strategy that relied on STS content mapping. The major motivation for constructing this map is to provide the directed genomic sequencing project at LBL with well- characterized clones from which to generate sequencing templates. Toward this goal we have established a production STS mapping effort at LBL capable of assigning more 100 STS markers per month to the physical map. The major source of STS markers for this project derive from the insert ends of P1 clones that have been positioned in the genome by in situ hybridization to polytene chromosomes. Other sources of STS markers being used for this project are known Drosophila genes and rescued plasmids derived from lethal P element insertions. As of 31 July 1994 we have mapped a total of 1848 STS markers from three classes: P1 end sequences, rescued P elements, and known Drosophila genes. A total of 1299 of these STS markers derive from P1 end sequences. Together these STS markers have assigned over 50% of the clones in the library to contigs, and these contigs cover about 70% of the euchromatin. At the end of the initial grant period (31 July 1995) we are confident that 90-95% of the euchromatic genome will be represented in P1 contigs. This section of the grant proposal is a renewal of this work and describes our approaches to map completion and contig closure. The experiments proposed in this section are organized in three sequential stages. First, the P1 clones remaining after the current production STS mapping phase exhausts the current supply of in situ localized P1 clones used to generate terminal STS markers will be assigned by additional STS mapping experiments. After all the clones in the library have been assigned to contigs, the second level of experiments proposed in this section are aimed at closing apparent gaps between contigs by additional STS mapping experiments. The final stage of map completion will be efforts to close gaps not represented in the master PI library. Closing these statistical gaps will be accomplished by screening additional P1 clones that provide another seven to eight genomic equivalents above and beyond the coverage provided by the master library. Clones to fill gaps remaining after these experiments will be sought by screening libraries constructed with other vector systems such as lambda, cosmid, and YAC.

最初的果蝇基因组中心拨款建议建立一个基于克隆的 黑腹果蝇常染色质基因组的物理图谱， 噬菌体P1文库和依赖于 STS内容映射。 绘制这幅地图的主要动机是 为LBL的定向基因组测序项目提供良好的- 从其产生测序模板的表征的克隆。 朝向 为了实现这一目标，我们在LBL建立了生产STS映射工作 能够每月为物理地图分配100个以上的STS标记。 本项目STS标记的主要来源来自插页 P1克隆的末端已经通过原位定位在基因组中， 与多线染色体杂交。 STS标记的其他来源是 用于这个项目的是已知的果蝇基因和拯救质粒 来自致命的P元素插入。 截至1994年7月31日， 共定位了来自三类的1848个STS标记：P1末端序列， 拯救的P元件和已知的果蝇基因。 其中，1299 STS标记来源于P1末端序列。 这些STS标记一起具有 将文库中超过50%的克隆分配给重叠群， 重叠群覆盖约70%的常染色质。 在最初的结尾， 补助金期间（1995年7月31日），我们有信心，90-95%的常色 基因组将以P1重叠群表示。 本节补助金 建议是这项工作的更新，并描述了我们的方法来映射 完成和重叠群封闭。 本节提出的实验 分为三个连续的阶段。 第一，P1克隆剩余 当前生产STS映射阶段耗尽当前供应后 用于产生末端STS标记的原位定位的P1克隆将 通过额外的STS映射实验分配。 在所有的克隆人 文库已分配到重叠群，第二级实验 本节中提出的方法旨在缩小重叠群之间的明显差距 通过额外的STS映射实验。 地图完成的最后阶段 将努力填补主PI库中没有的空白。 将通过筛选额外的 P1克隆提供上述另外七到八个基因组当量， 超出了主库的覆盖范围。 克隆以填补空白 这些实验后剩下的将通过筛选文库来寻找 用其它载体系统如λ、粘粒和YAC构建。