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从小鼠肝脏的膜和游离多核糖体制备的聚腺苷酸(poly(A))-mRNA的比较。

Comparison of poly(A)-mRNA prepared from membranes and free polyribosomes of mouse liver.

作者信息

Clissold P M, Mason P J, Bishop J O

出版信息

Proc Natl Acad Sci U S A. 1981 Jun;78(6):3697-701. doi: 10.1073/pnas.78.6.3697.

Abstract

We have compared poly(A)-mRNA isolated from mouse liver endoplasmic reticulum (ER) membranes (ER mRNA) with that from free polyribosomes (free mRNA) by translation in a cell-free system and by mRNA--cDNA reassociation analysis. The partitioning of certain translation products between the two fractions suggests that the physical contamination of the free mRNA by the ER is 10%, and contamination of the ER by the free is 1%. Reassociation crossreactions suggest that no sequences are totally absent from either fraction. In particular, the rare sequences are shared between the two fractions. Among the more abundant sequences, we detect three groups: (i) a group of about 10 sequences that are relatively abundant in both fractions; (ii) a group that occurs mostly in the free fraction; (iii) a group of about 12 sequences, 6 or 7 of them encoding secretory polypeptides. The members of this group are largely or entirely confined to the ER fraction. Secretory proteins are almost entirely absent from the products of th free mRNA fraction. Quantitation of individual mRNA sequences by hybridization with cDNA clones shows that a range of sequence concentrations exists within the abundant class of ER mRNA.

摘要

我们通过无细胞体系中的翻译以及mRNA-cDNA重缔合分析,对从小鼠肝脏内质网(ER)膜分离的聚腺苷酸(poly(A))-mRNA(ER mRNA)和游离多核糖体的聚腺苷酸-mRNA(游离mRNA)进行了比较。某些翻译产物在这两个组分之间的分配情况表明,游离mRNA受ER的物理污染为10%,而ER受游离mRNA的污染为1%。重缔合交叉反应表明,两个组分中均不存在完全缺失的序列。特别是,稀有序列在两个组分之间是共享的。在丰度较高的序列中,我们检测到三组:(i)一组约10个在两个组分中都相对丰富的序列;(ii)一组主要出现在游离组分中的序列;(iii)一组约12个序列,其中6或7个编码分泌性多肽。该组的成员在很大程度上或完全局限于ER组分。游离mRNA组分的产物中几乎完全不存在分泌蛋白。通过与cDNA克隆杂交对单个mRNA序列进行定量分析表明,在ER mRNA的丰富类别中存在一系列的序列浓度。

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