DETECTING AND MAPPING SEQUENCE CHANGES IN DNA

检测和绘制 DNA 序列变化

• 批准号: 3275739
• 负责人: ROSEMARY W ELLIOTT
• 金额: $ 17.75万
• 依托单位: ROSWELL PARK CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-12-01 至 1991-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3275739
• 关键词: DNA; autoradiography; biological polymorphism; chromosome complement; chromosomes; complementary DNA; endodeoxyribonuclease; genetic manipulation; genetic mapping; genetic markers; genetic strain; laboratory mouse; molecular cloning; nucleic acid hybridization; nucleic acid sequence; plasmids; radiotracer

The goal of the current study is the generation of a map of the mouse genome in which all chromosomes are marked by loci, spaced approximately ten map units apart. These loci should be characterized by polymorphic variation, such that any given pair of strains which do not have a recent inbreeding history have different alleles at about half the loci. The single type of genetic marker which could fulfill these requirements is the restriction fragment length polymorphism in mouse DNA. We plan to map these variants using a variety of mapping techniques. The main emphasis will be on use of recombinant inbred strains. Other techniques will involve somatic hybrids segregating mouse chromosomes, the classical backcross and congenic strains of mice. We plan to probe the DNA with several kinds of cloned sequences. First, we will use identified cloned cDNA probes obtained from other laboratories. These have intrinsic interest as well as adding to the map. Second, we will use probes which we know hybridize to chromosomes which have very few polymorphic loci. These clones will be identified after hybridizing to DNA from a panel of mouse-hamster hybrid cell lines. They will be of two types. We will use random cDNA clones from libraries from a number of mouse organs. We also plan to use flanking regions to endogenous retroviruses which we will clone for genomic libraries of DNA from C57BL mouse strains. We have shown that these retroviruses occupy loci in C57BL which are not allelic to retroviral loci in other strains. This implies that they will hybridize to polymorphic DNA fragments. The fragment will contain a retrovirus in C57BL and will lack a retrovirus in other strains. As retroviruses appear to insert randomly in the mouse genome and mouse strains each have about 130 copies, they will be spaced about 12 cM apart on the mouse genome, and will be present on every chromosome. The achievement of a fully marked mouse map is essential to the full interpretation and analysis of regulatory genes, multigene phenotypes, and tumor susceptibility.

当前研究的目标是生成小鼠的地图 基因组：所有染色体都由基因座标记的基因组，间隔约为 相隔十个地图单位 这些位点应具有多态性 变异，这样任何给定的一对菌株， 近亲繁殖历史在大约一半的基因座上具有不同的等位基因。 的 可以满足这些要求的单一类型的遗传标记是 限制性片段长度多态性。 我们计划绘制地图 这些变体使用各种映射技术。 主要的重点 将使用重组近交系。 其他技术将 涉及分离小鼠染色体的体细胞杂种， 回交和同类品系的小鼠。 我们计划用一种 几种克隆序列。 首先，我们将使用识别克隆 从其他实验室获得的cDNA探针。 这些具有内在的 #21453;感兴趣，并添加到地图上。 第二，我们将使用探针， 已知与具有很少多态位点的染色体杂交。 这些 克隆将在与来自一组DNA杂交后鉴定。 小鼠-仓鼠杂交细胞系。 他们将有两种类型。 我们将使用 随机的cDNA克隆，来自小鼠器官的文库。 我们也 我计划用侧翼区来克隆内源性逆转录病毒 用于C57 BL小鼠品系的DNA的基因组文库。 我们已经证明 这些逆转录病毒占据C57 BL中与逆转录病毒不等位的基因座 其他菌株中的基因座。 这意味着它们将与 多态性DNA片段 该片段将包含C57 BL中的逆转录病毒 并且在其他菌株中缺乏逆转录病毒。 因为逆转录病毒似乎 随机插入小鼠基因组中，小鼠品系各有约130个 拷贝，它们将在小鼠基因组上间隔约12 cM， 存在于每个染色体上。 一只完全标记的老鼠的成就 地图是必不可少的充分解释和分析的监管 基因、多基因表型和肿瘤易感性。