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牛促肾上腺皮质激素/β-促脂素前体编码信使核糖核酸的纯化与特性分析

Purification and characterization of the messenger RNA coding for bovine corticotropin/beta-lipotropin precursor.

作者信息

Kita T, Inoue A, Nakanishi S, Numa S

出版信息

Eur J Biochem. 1979 Jan 15;93(2):213-20. doi: 10.1111/j.1432-1033.1979.tb12813.x.

Abstract

The mRNA coding for the common precursor of corticotropin and beta-lipotropin has been purified to homogeneity from neurointermediate lobes of bovine pituitaries. The homogeneity of the mRNA preparation is evidenced by analysis of its translation product, electrophoresis on polyacrylamide gel in the presence of formamide and analysis of the kinetics of hybridization with its cDNA. The purification procedure involves the isolation of RNA from membrane-bound polysomes, chromatography on oligo(dT)-cellulose and on poly(U)-Sepharose and sucrose density gradient centrifugation. The mRNA has a molecular weight of approximately 450000, equivalent to approximately 1360 nucleotides in length, and contains a polyadenylate sequence with an average length of 68 nucleotides. The size of the mRNA is sufficiently large to encode the corticotropin/beta-lipotropin precursor.

摘要

编码促肾上腺皮质激素和β-促脂素共同前体的信使核糖核酸(mRNA)已从牛垂体神经中间叶中纯化至同质。通过对其翻译产物进行分析、在甲酰胺存在下于聚丙烯酰胺凝胶上进行电泳以及分析与互补脱氧核糖核酸(cDNA)杂交的动力学,证明了mRNA制剂的同质性。纯化过程包括从膜结合多核糖体中分离核糖核酸(RNA)、在寡聚(dT)-纤维素和聚(U)-琼脂糖凝胶上进行层析以及蔗糖密度梯度离心。该mRNA的分子量约为450000,相当于长度约为1360个核苷酸,并含有平均长度为68个核苷酸的聚腺苷酸序列。mRNA的大小足够大,能够编码促肾上腺皮质激素/β-促脂素前体。

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