RNA FINGERPRINTS--DIFFERENTIAL GENE EXPRESSION IN BRAIN

不同组织和发育过程中基因表达的差异 可以任意使用RNA快速有效地比较阶段 引物聚合酶链反应（RAP）指纹图谱方法， 在我们的实验室里开发的。 在RAP中，任意序列的引物 启动第一和第二链cDNA合成。 的混合物 产物经PCR扩增并经电泳解析， 具有组织特异性的高重现性指纹。 差异 指纹之间的差异来自差异表达的基因。 的 该方法对RNA进行采样，偏差很小，可以识别新的 差异调节转录本。 RAP技术最令人兴奋的应用之一是大脑 发展 理解空间和发展差异， 发育过程中的基因表达是理解 大脑的正常功能及其病理学。 RAP方法允许 同时比较多个RNA样品，因此， 能够在大量表达的基因样本中检测出 在大脑发育过程中只短暂表达， 只在大脑的一个或几个区域表达，或者是差异表达， 在正常大脑和神经退行性疾病患者大脑中的表达 紊乱 这些基因是有吸引力的候选者，可通过以下方法进行进一步分析： 常规战略。 我们将鉴定和描述这些基因 从老鼠的大脑中分离出来。 一个试点 研究已证明这个方法是可行的。

Differential gene expression between various tissues and developmental stages can be compared rapidly and efficiently using the RNA arbitrarily primed polymerase chain reaction (RAP) fingerprinting method that was developed in our laboratories. In RAP, a primer of arbitrary sequence primes both first and second strand cDNA synthesis. The mixture of products is PCR amplified and resolved electrophoretically, yielding highly reproducible fingerprints that are tissue-specific. Differences between fingerprints arise from differentially expressed genes. The method samples RNAs with little bias, allowing the identification of novel differentially regulated transcripts. One of the most exciting applications of RAP technology is to brain development. An understanding of spatial and developmental differences in gene expression during development is an important goal in understanding the normal function of the brain and its pathology. The RAP method allows the comparison of many RNA samples simultaneously and, therefore, is capable of detecting among a large sample of expressed genes those few that are expressed for only a brief period during brain development, are expressed in only one or a few areas of the brain, or are differentially expressed between normal brains and those with neurodegenerative disorders. Such genes are attractive candidates for further analysis by conventional strategies. We will identify and characterize such genes from the mouse brain in an international collaborative effort. A pilot study has already demonstrated the feasibility of this approach.