GENETIC MAPPING IN THE MOUSE BY PCR FINGERPRINTING

使用单个随机选择的引物的聚合酶链式反应(任意 引物聚合酶链式反应(Primed 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，这可能是 在重组近交系小鼠中进行遗传定位。这种方法是 非常快速和简单，只使用几毫微克的模板DNA。 引物可以单独使用，也可以成对使用。 组合。平均有四个以上的可映射的多态 BXD重组自交系中每对或每对引物的唯一性 台词。我们将能够使用少至35个聚合酶链式反应引物来放置 小鼠遗传图谱上另外的2400个DNA多态 利用26个BXD、25个AXB和25个BXA重组自交系。这个 这张地图的精确度应优于5厘米，并应 平均每一百万个碱基对中有一个多态性。这个 遗传分配的AP-PCR片段将是一种有价值的资源 多个限制性片段长度多态(RFLP)和聚合酶链式反应 基于已在地图上的(CA)n个多态。那些多态现象 与基因图谱突变共分离应该对 克隆导致这种突变的基因周围的区域。 例如，BXD、AXB和BXA重组自交系就是模式 对于某些疾病的易感性，如动脉粥样硬化， 糖皮质激素引起的腭裂、一些感染和一些肿瘤。 定位的AP-PCR多态也将有助于遗传 正在组装成一个小鼠基因组克隆的定位 重叠群图。此外，由于同步性，关于鼠标的信息 基因组有时可以延伸到人类身上，反之亦然。

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.