Savioz A, Davies R W
Robertson Laboratory of Biotechnology, University of Glasgow, UK.
Gene. 1995 Mar 10;154(2):225-30. doi: 10.1016/0378-1119(94)00864-o.
Many important phenomena of normal brain physiology and disease are likely to be related to the function of genes expressed in localised regions of the brain. We show that subtracted libraries enriched in clones corresponding to rare mRNAs, which must include genes with very localised and neuron-specific expression, can easily be produced from single-stranded directional cDNA libraries after hybridization to excess photobiotinylated opposite-stranded cDNA (or RNA) from another brain region, followed by the removal of biotinylated molecules. We also demonstrate the use of heterologous probes from anatomically precise small regions of bovine brain to identify cDNA clones that putatively represent mRNAs present at significantly higher levels in a substantia nigra mRNA population enriched for pars compacta mRNA than in the total ventral midbrain or cerebellar mRNA population. Some of these cDNAs may identify genes that play important roles in the specific molecular biology of dopaminergic neurons, including susceptibility to Parkinson's disease.
正常脑生理学和疾病的许多重要现象可能与大脑局部区域表达的基因功能有关。我们表明，在与来自另一个脑区的过量光生物素化的反义链cDNA（或RNA）杂交后，通过去除生物素化分子，能够轻松地从单链定向cDNA文库中产生富含与稀有mRNA对应的克隆的扣除文库，这些稀有mRNA必定包含具有非常局部化和神经元特异性表达的基因。我们还展示了使用来自牛脑解剖学精确小区域的异源探针来鉴定cDNA克隆，这些克隆可能代表在富含致密部mRNA的黑质mRNA群体中水平显著高于总腹侧中脑或小脑mRNA群体中的mRNA。其中一些cDNA可能鉴定出在多巴胺能神经元的特定分子生物学中起重要作用的基因，包括对帕金森病的易感性。
