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电鳐电运动神经元中两个同源cDNA克隆的分离与鉴定

Isolation and characterization of two homologous cDNA clones from Torpedo electromotor neurons.

作者信息

Ngsee J K, Scheller R H

机构信息

Department of Biological Sciences, Stanford University, CA 94305-5020.

出版信息

DNA. 1989 Oct;8(8):555-61. doi: 10.1089/dna.1989.8.555.

Abstract

Two homologous cDNA clones were isolated from a Torpedo california electric lobe lambda gt11 expression library using a polyclonal antiserum directed against proteins associated with synaptic vesicles. Northern blotting reveals an 8- to 9-kb transcript in the electric lobe and the spinal cord, but not in the brain or other non-neuronal tissues. Antibodies generated against a fusion protein synthesized in Escherichia coli reacted with a 85- to 90-kD species in the neurons of the electric lobe. The immunoreactivity is associated with microsomal membranes and can be extracted readily with high salt. Immunohistochemical studies demonstrated a sparse punctate staining pattern in the cell body which colocalized with a subpopulation of post-Golgi vesicles.

摘要

利用针对与突触小泡相关蛋白的多克隆抗血清,从加州电鳐的电叶λgt11表达文库中分离出两个同源cDNA克隆。Northern印迹分析显示,在电叶和脊髓中存在一个8至9kb的转录本,但在大脑或其他非神经组织中则没有。针对在大肠杆菌中合成的融合蛋白产生的抗体,与电叶神经元中一个85至90kD的蛋白发生反应。这种免疫反应性与微粒体膜相关,并且可以很容易地用高盐提取。免疫组织化学研究表明,在细胞体中存在稀疏的点状染色模式,该模式与高尔基体后小泡的一个亚群共定位。

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