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赠款提议建立一个基于克隆的 果蝇的物理图使用A 噬菌体P1库和依赖的非随机映射策略 STS内容映射。 构建此地图的主要动机是 提供LBL的定向基因组测序项目 表征从中生成测序模板的克隆。 走向 这个目标我们已经在LBL建立了生产STS映射工作 能够为物理地图分配更多100个STS标记。 该项目的STS标记的主要来源是源自插入物 原位位于基因组中的P1克隆的末端 与聚单抗染色体的杂交。 STS标记的其他来源是 该项目用于该项目是已知的果蝇基因和救出的质粒 源自致命的P元素插入。 截至1994年7月31日，我们有 从三个类别绘制了总共1848个STS标记：P1结束序列， 营救了P元素和已知的果蝇基因。 总共1299 STS标记源自P1末端序列。 这些STS标记在一起 将图书馆中50％以上的克隆分配给重叠群，这些 重叠群覆盖大约70％的白染色质。 在初始结束时 赠款期（1995年7月31日），我们相信90-95％ 基因组将在P1重叠群中表示。 赠款的这一部分 提案是对这项工作的续签，并描述了我们的地图的方法 完成和关闭。 本节提出的实验 分为三个顺序阶段。 首先，剩余的P1克隆 当前生产STS映射阶段耗尽当前电源 用于生成终端STS标记的原位局部P1克隆的 通过其他STS映射实验分配。 在所有克隆之后 库已分配给重叠群，第二级实验 本节中提出的旨在缩小重叠之间的明显差距 通过其他STS映射实验。 地图完成的最后阶段 将努力弥合主PI库中未代表的差距。 结束这些统计差距将通过筛选额外来完成 P1克隆，在上方提供了另外七到八个基因组当量的克隆和 超越大师库提供的覆盖范围。 克隆以填补空白 在这些实验之后剩下的将通过筛选图书馆寻求 由Lambda，Cosmid和YAC等其他向量系统构建。