从正常大脑和阿尔茨海默病患者大脑中克隆cDNA至表达载体λGT 11。

Cloning of the cDNA from normal brain and brain of patients with Alzheimer's disease in the expression vector lambda GT 11.

Octave J N, de Sauvage F, Macq A F, Maloteaux J M

Université Catholique de Louvain, Laboratoire de Neurochimie, Brussels, Belgium.

Prog Neuropsychopharmacol Biol Psychiatry. 1988;12(5):813-20. doi: 10.1016/0278-5846(88)90026-7.

Abstract

RNA was purified from postmortem human brains, and the poly A+ RNA was isolated by oligo dT cellulose. 2. Double stranded cDNA was synthesized using reverse transcriptase, RNAse H and DNA polymerase. 3. cDNA was cloned in the lambda GT 11 expression vector, and libraries containing between 1 and 2 millions clones were obtained. 92 to 98% of the plaques contained a recombinant phage. 4. Such libraries will allow the molecular characterization of cDNA and corresponding proteins which play a key role in brain functions and in particular which could be involved in the etiology of Alzheimer's dementia.

摘要

从人死后的大脑中纯化RNA，并用寡聚dT纤维素分离出聚腺苷酸加尾RNA。2. 使用逆转录酶、核糖核酸酶H和DNA聚合酶合成双链cDNA。3. 将cDNA克隆到λGT 11表达载体中，获得了包含100万至200万个克隆的文库。92%至98%的噬菌斑含有重组噬菌体。4. 这样的文库将有助于对在脑功能中起关键作用、特别是可能与阿尔茨海默病痴呆病因有关的cDNA和相应蛋白质进行分子表征。