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[小鼠浆细胞瘤的信使核糖核酸。无细胞系统中的逆转录与翻译]

[mRNA of mouse plasmacytoma. Reverse transcription and translation in cell-free systems].

作者信息

Sakharova N K, Ryndich A V, Kavsan V M, Goriunova L E, Grechko V V

出版信息

Mol Biol (Mosk). 1979 Jan-Feb;13(1):169-79.

PMID:88667
Abstract

Total poly(A)-mRNA from polyribosomes of MOPC 21 mouse myeloma were investigated. Poly(A)-mRNA was released by two successive chromatography on oligo (dT)-cellulose. A 14S fraction of total poly(A)-mRNA was obtained and partially purified by sucrose gradient centrifigation followed by acrylamide gel electrophoresis. As estimated from the electrophoretic analysis, the 14S mRNA has three components, one of which appears to be 18S rRNA and two others--mRNAs with molecular weight of 5.2.10(5) and 3.8.10(5), respectively. Total poly(A)-mRNA and partially purified 14S mRNA were active when employed as a template in a reverse transcription and cell-free system from wheat germ. DNA complementary to the 14S mRNA was prepared with avian myeloblastosis virus RNA-dependent DNA polymerase. This cDNA was heterogeneous in size with the average size of about 800 nucleotides when analyzed by gel electrophoresis in 98% formamide. The maximal length was about 1100 nucleotides that consistent with full template length. About half of the translation product directed by the 14S mRNA migrated as mature L-chain Ig (upon polyacrylamide gel electrophoresis in sodium dodecylsulfate). The presented data suggested that 14S mRNA species contain mRNA L-chain Ig.

摘要

对MOPC 21小鼠骨髓瘤多核糖体中的总聚腺苷酸(poly(A))-mRNA进行了研究。通过在寡聚(dT)-纤维素上连续两次层析释放出poly(A)-mRNA。获得了总poly(A)-mRNA的14S组分,并通过蔗糖梯度离心随后进行丙烯酰胺凝胶电泳进行部分纯化。根据电泳分析估计,14S mRNA有三个组分,其中一个似乎是18S rRNA,另外两个分别是分子量为5.2×10⁵和3.8×10⁵的mRNA。当总poly(A)-mRNA和部分纯化的14S mRNA在小麦胚芽的反转录和无细胞系统中用作模板时具有活性。用禽成髓细胞瘤病毒RNA依赖性DNA聚合酶制备了与14S mRNA互补的DNA。当在98%甲酰胺中通过凝胶电泳分析时,该cDNA大小不均一,平均大小约为800个核苷酸。最大长度约为1100个核苷酸,与完整模板长度一致。由14S mRNA指导的约一半翻译产物在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中迁移为成熟的轻链Ig。所呈现的数据表明14S mRNA种类包含轻链Ig的mRNA。

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