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一种利用翻译释放的mRNA进行cDNA克隆的直接方法。

A direct method for cDNA cloning employing translation-released mRNA.

作者信息

Barardi C R, Santos C L, Villa L L, Brentani R R

机构信息

Instituto Ludwig de Pesquisa sobre o Câncer, São Paulo, Brasil.

出版信息

Braz J Med Biol Res. 1987;20(3-4):313-20.

PMID:3451785
Abstract
  1. A new cloning procedure is described for cDNA synthesis from mRNA released by in vitro translation of polysomes in a cell-free amino acid incorporating system. The usefulness of the method lies in the feasibility of employing nanogram amounts of mRNA. 2. Complementary DNA is synthesized directly in the translation mixture simply by adjusting the concentration of some components and removing ribosomes by boiling and centrifugation. 3. As an example, we report here the construction and characterization of a cDNA clone corresponding to chick alpha(I)procollagen starting from a collagen-synthesizing polysome fraction obtained from chick embryos.
摘要
  1. 本文描述了一种新的克隆程序,用于在无细胞氨基酸掺入系统中,从多核糖体体外翻译释放的mRNA合成cDNA。该方法的实用性在于使用纳克量mRNA的可行性。2. 通过简单地调整某些成分的浓度并通过煮沸和离心去除核糖体,可直接在翻译混合物中合成互补DNA。3. 作为一个例子,我们在此报告从鸡胚获得的胶原合成多核糖体组分开始构建和鉴定与鸡α(I)前胶原相对应的cDNA克隆。

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