从富含多核糖体的组分中纯化和鉴定前胶原信使核糖核酸

Purification and characterization of procollagen mRNA from a heavy polysomal fraction.

作者信息

dos-Santos C L, Hatada E H, D'Abronzo F, Sonohara S, Brentani R R

出版信息

Braz J Med Biol Res. 1982 Dec;15(6):349-54.

Abstract

A method is described for the purification of intact procollagen mRNA from heavy polysomal fractions isolated from 10 day-old chick embryos. Procollagen mRNA activity was characterized by gel electrophoresis under denaturing conditions and by its activity in in vitro cell-free translation systems derived from wheat germ and rabbit reticulocytes. 7-Methyl-guanosine monophosphate inhibited the translation of procollagen mRNA and globin mRNA to the same extent. Collagenase-sensitive polypeptides synthesized displayed the same electrophoretical mobilities as marker procollagen chains. The purity of the mRNA was 78% on the basis of ROt plot analysis of hybridization with its corresponding cDNA, using globin cDNA as the reference for 100%.

摘要

本文描述了一种从10日龄鸡胚分离的重多核糖体组分中纯化完整前胶原mRNA的方法。前胶原mRNA活性通过变性条件下的凝胶电泳及其在源自小麦胚芽和兔网织红细胞的体外无细胞翻译系统中的活性来表征。7-甲基鸟苷单磷酸对前胶原mRNA和珠蛋白mRNA的翻译抑制程度相同。合成的胶原酶敏感多肽显示出与标记前胶原链相同的电泳迁移率。以珠蛋白cDNA作为100%的参考，根据与其相应cDNA杂交的Rot图分析，mRNA的纯度为78%。