大鼠脑谷氨酸脱羧酶信使核糖核酸的分子克隆、表达及原位杂交
Molecular cloning, expression and in situ hybridization of rat brain glutamic acid decarboxylase messenger RNA.
作者信息
Julien J F, Legay F, Dumas S, Tappaz M, Mallet J
出版信息
Neurosci Lett. 1987 Jan 14;73(2):173-80. doi: 10.1016/0304-3940(87)90013-9.
A cDNA library was generated in the expression vector lambda GT11 from rat brain poly(A)+ RNAs and screened with a GAD antiserum. Two clones reacted positively. One of them was shown to express a GAD activity which was specifically trapped on anti-GAD immunogel and was inhibited by gamma-acetylenic-GABA. Blot hybridization analysis of RNAs from rat brain revealed a single 4 kilobases band. Preliminary in situ hybridizations showed numerous cells labelled by the GAD probe such as the Purkinje and stellate cells in the cerebellar cortex and the cells of the reticular thalamic nucleus.
利用大鼠脑多聚腺苷酸（poly(A)）+RNA 在表达载体λGT11 中构建了一个 cDNA 文库，并用谷氨酸脱羧酶（GAD）抗血清进行筛选。有两个克隆呈阳性反应。其中一个克隆显示表达一种 GAD 活性，该活性可特异性地被抗 GAD 免疫凝胶捕获，并被γ-炔丙基-GABA 抑制。对大鼠脑 RNA 的印迹杂交分析显示有一条单一的 4 千碱基条带。初步原位杂交显示，许多细胞被 GAD 探针标记，如小脑皮质中的浦肯野细胞和星状细胞以及丘脑网状核的细胞。
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