GENETIC MAPPING IN THE MOUSE BY PCR FINGERPRINTING

通过 PCR 指纹图谱绘制小鼠基因图谱

• 批准号: 3333633
• 负责人: MICHAEL MCCLELLAND
• 金额: $ 23.57万
• 依托单位: CALIFORNIA INSTITUTE OF BIOLOGICAL RES
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-03-15 至 1995-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3333633
• 关键词: DNA footprinting; gene expression; gene mutation; genetic mapping; genetic markers; genetic polymorphism; genome; laboratory mouse; molecular cloning; phenotype; polymerase chain reaction; restriction fragment length polymorphism

A PCR method employing single arbitrarily chosen primers (Arbitrarily primed PCR) generates consistent 'fingerprints' for the mouse genome. These fingerprints include DNA fragment polymorphisms which can be genetically mapped in recombinant inbred lines of mice. This method is very fast and simple and uses only a few nanograms of template DNA. Primers can be used individually or in almost every pairwise combination. There are an average of over four mappable polymorphisms unique to each primer or pair of primers in the BXD recombinant inbred lines. We will be able to use as few as 35 PCR primers to place an additional 2400 DNA polymorphisms on the mouse genetic map in three years using the 26 BXD, 25 AXB and 25 BXA recombinant inbred lines. The accuracy of this map should be better than 5 centiMorgans and should have an average of almost one polymorphism per million base pairs. The genetically assigned AP-PCR fragments will be a valuable resource along with the many restriction fragment length polymorphisms (RFLPs) and PCR based (CA)n polymorphisms already on the map. Those polymorphisms that co-segregate with genetically mapped mutations should be useful for cloning the regions around the genes responsible for such mutations. For instance, the BXD, AXB and BXA recombinant inbred lines are models for susceptibility to certain diseases such as atherosclerosis, glucocorticoid induced cleft palate, some infections and some neoplasms. Mapped AP-PCR polymorphisms will also facilitate the genetic localization of mouse genomic clones that are being assembled into a contig map. Furthermore, due to synteny, information about the mouse genome can sometimes be extended to humans and vice versa.

使用单个任意选择的引物（顺序地）的PCR方法 引物PCR）为小鼠基因组产生一致的“指纹”。 这些指纹包括DNA片段多态性， 在小鼠的重组近交系中进行遗传定位。 该方法 非常快速和简单，只需要几纳克的模板DNA。 引物可以单独使用，也可以几乎成对使用 组合. 平均有四个以上的可作图多态性 对于BXD重组近交系中的每个引物或引物对是唯一的 线 我们将能够使用少至35个PCR引物， 另外2400 DNA多态性的小鼠遗传图谱在三个 利用26个BXD、25个AXB和25个BXA重组自交系， 的 该地图的精度应优于5厘米摩根，并应 平均每一百万个碱基对就有一个多态性。 的 基因分配的AP-PCR片段将是沿着 许多限制性片段长度多态性（RFLP）和PCR 基于（CA）n多态性已经在地图上。 这些多态性 与基因定位突变共分离应该是有用的， 克隆导致这种突变的基因周围的区域。 例如，BXD、AXB和BXA重组近交系是模型 对于某些疾病如动脉粥样硬化的易感性， 糖皮质激素引起的腭裂，一些感染和一些肿瘤。 作图的AP-PCR多态性也将促进遗传 小鼠基因组克隆的定位， 重叠群图。 此外，由于共线性，关于小鼠的信息 基因组有时可以扩展到人类，反之亦然。